Using bacteria to make a biomolecule Using bacteria to make a biomolecule

Introduction “It had been known for some years that treatment of cells in culture with one virus (the interfering virus) blocked the growth Bacteria are single-celled organisms with a relatively simple of a second virus, called the challenge virus... However, the biochemistry. Advances in genetic engineering and mechanism of virus interference was completely unknown.” recombinant DNA technology have made it possible to use them for the mass-production of useful biomolecules such as D. Burke, on the discovery of interferon by Alick Isaacs and Jean Lindenmann hormones, vitamins and medicines. Many of these compounds were formerly sourced from animals. Obtaining pure was used to synthesise complementary DNA (cDNA). The cDNA was MSD Medical Education compounds in sufficient quantities was very difficult and isolated and then added to mRNA from the non-induced culture. cDNA that expensive. Animal-derived biochemicals might not be did not associate with mRNA could be taken as being involved with MSD works with leading medical education partners to deliver completely effective in humans, an environment they were not interferon synthesis. Having examined and sequenced this DNA, scientists ongoing medical education training and was the first company to partner with the BMJ to bring their highly regarded made for. Adverse immune reactions could, and did, happen. could now look for, and obtain from genomes, DNA coding for interferon. BMJ Masterclasses to Ireland. This DNA was introduced into E.coli bacteria which multiplied and produced commercial quantities of interferon. The cells were regularly harvested and The renowned Merck Manuals, which have set the standard for About interferon the interferon extracted. Genetic modification of the bacteria was necessary excellence in professional medical information since they were as otherwise they would destroy interferon, a protein foreign to them. first published in 1899 are also accessible via the univadis portal. The story of interferon began in the 1950s. Scientists were looking for Insertion of the DNA into E.coli requires first its insertion intoplasmids or Fig. 2. Electron micrograph of E. coli This includes one of the world’s most widely used medical antibiotics that would combat viruses at a time when many vaccines had textbooks, The Merck Manual of Diagnosis and Therapy. yet to be developed. In 1957, Lindenmann and Isaacs inoculated chicken phages (viruses that infect bacteria) which are used as cloning vectors. Enzymes are used to excise DNA from plasmids and to insert interferon- embryos with influenza virus and noted that, after a time, viral action was Millions of people around the world are living longer, more These vectors are brought into contact with bacterial cells in the hope that coding DNA enzymatically obtained from human cells. Other DNA inhibited. The tissues were cleared of virus but, if influenza or other viruses productive lives, thanks to improvements in healthcare, including they will carry interferon DNA into the cells. Chemicals may be added to sequences, such as those coding for glycosylation, are also inserted. were subsequently applied, the anti-viral property was found to have access to innovative medicines and vaccines. facilitate their interaction. Plasmids are circular DNA strands which can be E.coli are prokaryotic cells, lacking some of the machinery required for persisted. Interferon had been discovered. passed between bacteria. They replicate with the bacteria and can also processing proteins characteristically produced by eukaryotic cells. At MSD in Ireland we are playing our part, working closely with replicate autonomously. (They are implicated in the transfer of antibiotic Time would show that there were many types of interferon. They are In E.coli, genes for abundantly expressed proteins have a low percentage many Irish charities, not-for-profit organisations and centres of resistance). cytokines - types of glycoprotein, that are produced by cells in response of particular codons so, before insertion, foreign DNA may need to be academic excellence to improve access to healthcare. Our to the presence of viruses and certain foreign chemicals. Their action When viruses enter cells, they may incorporate DNA into the host DNA modified to reflect this. Interferon genes do not haveintrons (non-coding current areas of focus include research partnerships, medical is indirect, activating genes in neighbouring cells to make proteins that (lysogenic cycle). They may also replicate using the host cell’s resources sequences) which might otherwise complicate their operation in the bacterial grants, patient access programmes and support of patient groups in their advocacy, policy and awareness work. interfere with cell replication. Apoptosis in infected cells may with the release of virus particles (lytic cycle). Which cycle a virus enters cell (Enzymes would have been required to remove the non-coding stretches be encouraged. depends on environmental conditions and on the type of virus involved. from the mRNA initially formed). It is possible to produce synthetic genes and this can refine the whole process. There have also been experiments MSD Public Health Education Interferon mobilises cells of the immune system and can cause foreign cells Plasmids and phages are natural phenomena that provide a means for transmitting DNA into bacteria and other cells. on the production of hybrid interferons. (including cancer cells) to display MHC antigens (major histocompatibility The ability of patients to read and understand healthcare complex) – making them easier targets for the body’s defences. A marker gene for resistance to an antibiotic can be inserted with the information is vital to ongoing and effective health management, Interferons-α and β are produced by all cells. Interferon-γ is produced by interferon gene. The bacteria replicate and are then treated with the however that is not always as straight forward as it might seem. antibiotic. A large percentage of bacteria fail to incorporate the foreign DNA Research suggests that people with low health literacy make immune cells only, stimulating further immune action. It also stimulates Bacterial DNA Plasmids expression by professional antigen-presenting cells; these cells kill – this procedure destroys them, leaving a clone of interferon-producing more mistakes with medication or treatment, are less able to follow treatment instructions, have difficulties negotiating the foreign cells and display antigens from their victims on their own surfaces, organisms. Further chemical processing of the interferon may be necessary healthcare system, and are more likely to be hospitalised than enhancing the immune reaction. to make it therapeutically effective. Polyethylene glycol may be combined people with adequate health literacy. We recognise that as a with it. “Pegylated interferon” breaks down more slowly in the human body, leader in healthcare we have a role to play in helping address this Interferons have anti-cancer properties, unsurprising as they inhibit cell necessitating fewer injections. proliferation. They can sometimes disrupt the enzyme action involved in vitally important issue. tumour angiogenesis (the laying on of a blood supply). It must be stated Phage vectors, like plasmids, are subjected to genetic modification, with In 2007, we formed a partnership with the National Adult Literacy that viruses and tumours have shown adaptations to interferon action. DNA being added and removed – their action has been effective. Gene Agency (NALA) with the specific aim of increasing public Tumours may employ different enzymes. Viruses may disrupt interferon therapy, where virus vectors are used to insert interferon genes directly into awareness of the health literacy issue, and to engage with cells in living people, is being researched and may become more important. receptors as well as producing chemicals to neutralise inhibitors of their healthcare professionals. Over the past six years this partnership Yeasts have been genetically engineered to produce interferon, an replication process. Nevertheless, interferons are effective overall. They also has helped Ireland become an internationally recognised centre have anti-bacterial properties and provide defence against other parasites. advantage being that they are eukaryotic cells. Other scientists have of excellence in health literacy. The highly successful annual successfully used virus vectors to produce interferon in insect cell cultures. Crystal Clear MSD Health Literacy Awards is just one high profile element amongst a number of initiatives. MSD also provides Production Cell information to the public on a wide range of health topics in Final words collaboration with partner organisations and through health Plasmid integration replication Only minute quantities are produced by cells. Research indicated that education programmes in such areas as women’s health, interferons were not effective across species although this is not always the Interferon was originally hailed as the wonder drug that would conquer hepatitis C, HIV, diabetes, osteoporosis and gastroenterology. case. Scientists made human cell cultures produce it by inducing them with cancer and other diseases. This was an inflated expectation. However, it viruses or certain chemicals. The protein was extracted and purified using is a useful medicine which biotechnology has made widely available. It is For more information visit www.msd-ireland.com different techniques including chromatography and monoclonal used in the treatment of some cancers (Kaposi’s sarcoma, leukemias), antibodies and its structure was determined. chronic hepatitis B and C, genital warts, multiple sclerosis and other conditions. It has side effects (generally controllable) which include flu-like In one method of obtaining interferon DNA, researchers used divided Integrated plasmid symptoms, lethargy, digestive upset, nosebleeds and perhaps immune Find this and other lessons on www.sta.ie identical tissue cultures. Interferon production was induced in one of them. depression. The overall actions of the interferons are far from fully mRNA was obtained from both cultures. mRNA from the induced culture Fig. 1. Two ways in which plasmids may be replicated in bacterial cells understood but their study and use are set to continue. Using bacteria to make a biomolecule Using bacteria to make a biomolecule

Introduction “It had been known for some years that treatment of cells in culture with one virus (the interfering virus) blocked the growth Bacteria are single-celled organisms with a relatively simple of a second virus, called the challenge virus... However, the biochemistry. Advances in genetic engineering and mechanism of virus interference was completely unknown.” recombinant DNA technology have made it possible to use them for the mass-production of useful biomolecules such as D. Burke, on the discovery of interferon by Alick Isaacs and Jean Lindenmann hormones, vitamins and medicines. Many of these compounds were formerly sourced from animals. Obtaining pure was used to synthesise complementary DNA (cDNA). The cDNA was MSD Medical Education compounds in sufficient quantities was very difficult and isolated and then added to mRNA from the non-induced culture. cDNA that expensive. Animal-derived biochemicals might not be did not associate with mRNA could be taken as being involved with MSD works with leading medical education partners to deliver completely effective in humans, an environment they were not interferon synthesis. Having examined and sequenced this DNA, scientists ongoing medical education training and was the first company to partner with the BMJ to bring their highly regarded made for. Adverse immune reactions could, and did, happen. could now look for, and obtain from genomes, DNA coding for interferon. BMJ Masterclasses to Ireland. This DNA was introduced into E.coli bacteria which multiplied and produced commercial quantities of interferon. The cells were regularly harvested and The renowned Merck Manuals, which have set the standard for About interferon the interferon extracted. Genetic modification of the bacteria was necessary excellence in professional medical information since they were as otherwise they would destroy interferon, a protein foreign to them. first published in 1899 are also accessible via the univadis portal. The story of interferon began in the 1950s. Scientists were looking for Insertion of the DNA into E.coli requires first its insertion intoplasmids or Fig. 2. Electron micrograph of E. coli This includes one of the world’s most widely used medical antibiotics that would combat viruses at a time when many vaccines had textbooks, The Merck Manual of Diagnosis and Therapy. yet to be developed. In 1957, Lindenmann and Isaacs inoculated chicken phages (viruses that infect bacteria) which are used as cloning vectors. Enzymes are used to excise DNA from plasmids and to insert interferon- embryos with influenza virus and noted that, after a time, viral action was Millions of people around the world are living longer, more These vectors are brought into contact with bacterial cells in the hope that coding DNA enzymatically obtained from human cells. Other DNA inhibited. The tissues were cleared of virus but, if influenza or other viruses productive lives, thanks to improvements in healthcare, including they will carry interferon DNA into the cells. Chemicals may be added to sequences, such as those coding for glycosylation, are also inserted. were subsequently applied, the anti-viral property was found to have access to innovative medicines and vaccines. facilitate their interaction. Plasmids are circular DNA strands which can be E.coli are prokaryotic cells, lacking some of the machinery required for persisted. Interferon had been discovered. passed between bacteria. They replicate with the bacteria and can also processing proteins characteristically produced by eukaryotic cells. At MSD in Ireland we are playing our part, working closely with replicate autonomously. (They are implicated in the transfer of antibiotic Time would show that there were many types of interferon. They are In E.coli, genes for abundantly expressed proteins have a low percentage many Irish charities, not-for-profit organisations and centres of resistance). cytokines - types of glycoprotein, that are produced by cells in response of particular codons so, before insertion, foreign DNA may need to be academic excellence to improve access to healthcare. Our to the presence of viruses and certain foreign chemicals. Their action When viruses enter cells, they may incorporate DNA into the host DNA modified to reflect this. Interferon genes do not have introns (non-coding current areas of focus include research partnerships, medical is indirect, activating genes in neighbouring cells to make proteins that (lysogenic cycle). They may also replicate using the host cell’s resources sequences) which might otherwise complicate their operation in the bacterial grants, patient access programmes and support of patient groups in their advocacy, policy and awareness work. interfere with cell replication. Apoptosis in infected cells may with the release of virus particles (lytic cycle). Which cycle a virus enters cell (Enzymes would have been required to remove the non-coding stretches be encouraged. depends on environmental conditions and on the type of virus involved. from the mRNA initially formed). It is possible to produce synthetic genes and this can refine the whole process. There have also been experiments MSD Public Health Education Interferon mobilises cells of the immune system and can cause foreign cells Plasmids and phages are natural phenomena that provide a means for transmitting DNA into bacteria and other cells. on the production of hybrid interferons. (including cancer cells) to display MHC antigens (major histocompatibility The ability of patients to read and understand healthcare complex) – making them easier targets for the body’s defences. A marker gene for resistance to an antibiotic can be inserted with the information is vital to ongoing and effective health management, Interferons-α and β are produced by all cells. Interferon-γ is produced by interferon gene. The bacteria replicate and are then treated with the however that is not always as straight forward as it might seem. antibiotic. A large percentage of bacteria fail to incorporate the foreign DNA Research suggests that people with low health literacy make immune cells only, stimulating further immune action. It also stimulates Bacterial DNA Plasmids expression by professional antigen-presenting cells; these cells kill – this procedure destroys them, leaving a clone of interferon-producing more mistakes with medication or treatment, are less able to follow treatment instructions, have difficulties negotiating the foreign cells and display antigens from their victims on their own surfaces, organisms. Further chemical processing of the interferon may be necessary healthcare system, and are more likely to be hospitalised than enhancing the immune reaction. to make it therapeutically effective. Polyethylene glycol may be combined people with adequate health literacy. We recognise that as a with it. “Pegylated interferon” breaks down more slowly in the human body, leader in healthcare we have a role to play in helping address this Interferons have anti-cancer properties, unsurprising as they inhibit cell necessitating fewer injections. proliferation. They can sometimes disrupt the enzyme action involved in vitally important issue. tumour angiogenesis (the laying on of a blood supply). It must be stated Phage vectors, like plasmids, are subjected to genetic modification, with In 2007, we formed a partnership with the National Adult Literacy that viruses and tumours have shown adaptations to interferon action. DNA being added and removed – their action has been effective. Gene Agency (NALA) with the specific aim of increasing public Tumours may employ different enzymes. Viruses may disrupt interferon therapy, where virus vectors are used to insert interferon genes directly into awareness of the health literacy issue, and to engage with cells in living people, is being researched and may become more important. receptors as well as producing chemicals to neutralise inhibitors of their healthcare professionals. Over the past six years this partnership Yeasts have been genetically engineered to produce interferon, an replication process. Nevertheless, interferons are effective overall. They also has helped Ireland become an internationally recognised centre have anti-bacterial properties and provide defence against other parasites. advantage being that they are eukaryotic cells. Other scientists have of excellence in health literacy. The highly successful annual successfully used virus vectors to produce interferon in insect cell cultures. Crystal Clear MSD Health Literacy Awards is just one high profile element amongst a number of initiatives. MSD also provides Production Cell information to the public on a wide range of health topics in Final words collaboration with partner organisations and through health Plasmid integration replication Only minute quantities are produced by cells. Research indicated that education programmes in such areas as women’s health, interferons were not effective across species although this is not always the Interferon was originally hailed as the wonder drug that would conquer hepatitis C, HIV, diabetes, osteoporosis and gastroenterology. case. Scientists made human cell cultures produce it by inducing them with cancer and other diseases. This was an inflated expectation. However, it viruses or certain chemicals. The protein was extracted and purified using is a useful medicine which biotechnology has made widely available. It is For more information visit www.msd-ireland.com different techniques including chromatography and monoclonal used in the treatment of some cancers (Kaposi’s sarcoma, leukemias), antibodies and its structure was determined. chronic hepatitis B and C, genital warts, multiple sclerosis and other conditions. It has side effects (generally controllable) which include flu-like In one method of obtaining interferon DNA, researchers used divided Integrated plasmid symptoms, lethargy, digestive upset, nosebleeds and perhaps immune Find this and other lessons on www.sta.ie identical tissue cultures. Interferon production was induced in one of them. depression. The overall actions of the interferons are far from fully mRNA was obtained from both cultures. mRNA from the induced culture Fig. 1. Two ways in which plasmids may be replicated in bacterial cells understood but their study and use are set to continue. UsingUsing bacteria bacteria to to makemake a a biomolecule biomolecule Using bacteria to make a biomolecule

SyllabusSyllabus references references StudentStudent Activities Activities ExaminationExamination Questions Questions DidDid You You Know? Know?

TheThe main main syllabus syllabus references references for for the the lesson lesson are: are: 1.1. Make Make a alist list of of the the different different human human interferons. interferons. LeavingLeaving Certificate Certificate Biology Biology 2013 2013 (HL) (HL) Q. Q.12 12 • • Interferon-γInterferon-γ has has been been used used effectively effectively in in the the treatment treatment of of osteopetrosis,osteopetrosis, an an inherited inherited disorder disorder characterised characterised by by an an overgrowth overgrowth JuniorJunior Certificate Certificate Science Science 2.2. Draw Draw a aposter poster picture picture of of an an interferon interferon molecule. molecule. (a)(a) A Avirus virus has has been been described described as as a apiece piece of of genetic genetic material material that that has has ofof bone bone which which leads leads to to the the development development of of deafness deafness and and blindness. blindness. 1C8.1C8. MicrobiologyMicrobiology and and biotechnology biotechnology escapedescaped from from a acell. cell. Give Give one one piece piece of of evidence evidence that that supports supports this this ThereThere is isreduced reduced space space for for bone bone marrow marrow resulting resulting in in anaemia anaemia and and 3.3. Draw Draw a apicture picture narrative narrative illustrating illustrating the the manufacture manufacture of of interferon, interferon, description.description. impairedimpaired resistance resistance to to disease. disease. fromfrom the the isolation isolation of of the the required required DNA DNA to to putting putting interferon interferon powder powder LeavingLeaving Certificate Certificate Biology Biology intointo a abottle. bottle. VirusesViruses are are examples examples of of obligate obligate parasites. parasites. Explain Explain why why this this is isthe the • • MultipleMultiple sclerosis sclerosis is isan an auto-immune auto-immune disorder disorder where where the the body’s body’s defencesdefences attack attack the the myelin myelin sheath sheath that that encloses encloses neurons. neurons. Impaired Impaired 2.1.4.2.1.4. ProkaryoticProkaryotic and and eukaryotic eukaryotic cells cells case.case. Give Give an an example example of of how how a avirus virus might might be be beneficial beneficial to to 4.4. Why Why might might it itbe be a agood good idea idea to to give give interferon interferon by by injection injection rather rather mankind.mankind. nervenerve impulse impulse transmission transmission follows, follows, leading leading to to weakness weakness and and 2.52.5 GeneticsGenetics thanthan orally? orally? paralysis.paralysis. Interferon Interferon therapy therapy appears appears to to work work partly partly by by slowing slowing 3.1.1.3.1.1. DiversityDiversity of of organisms organisms (b)(b) NameName the the kingdom kingdom to to which which bacteria bacteria belong. belong. Draw Draw a alarge large diagram diagram downdown the the production production of of T-cells T-cells 5.5. E.coli E.coli bacteria bacteria can can make make interferon. interferon. So So can can yeast yeast cells. cells. There There is is ofof a abacterial bacterial cell cell to to show: show: • • DiphtheriaDiphtheria and and botulism botulism toxins toxins are are produced produced by by genes genes inserted inserted by by 3.1.2.3.1.2. Micro-organismsMicro-organisms alsoalso experimentation experimentation with with cell-free cell-free methods. methods. Can Can you you think think of of 1.1. The The relative relative positions positions of of the the cell cell wall, wall, cell cell membrane membrane and and virusesviruses into into bacteria. bacteria. Scientists Scientists have have also also discovered discovered that that genes genes 3.1.3.3.1.3. Monera,Monera, e.g. e.g. bacteria bacteria advantages/disadvantagesadvantages/disadvantages associated associated with with each each method? method? capsule.capsule. 2. 2. A Aplasmid. plasmid. veryvery similar similar to to those those coding coding for for some some fusion fusion proteins proteins in in human human cells cells 3.1.9.3.1.9. NatureNature of of bacteria bacteria and and fungi. fungi. areare also also found found in in viruses. viruses. These These proteins proteins enable enable cells cells to to organise organise 6.6. Plasmids Plasmids and and phages phages are are vectors vectors for for introducing introducing DNA DNA into into bacterial, bacterial, LabelLabel each each of of the the above above structures. structures. andand other, other, cells. cells. The The future future may may see see increased increased use use of of cosmids cosmids and and intointo tissues tissues and and organs organs whereas whereas the the virus virus uses uses them them to to gain gain access access ScienceScience and and Technology Technology in in Action Action is isalso also widely widely used used artificialartificial chromosomes. chromosomes. WhatWhat are are the the drawbacks drawbacks and and prospects prospects UnderUnder what what circumstances circumstances does does a abacterial bacterial cell cell form form an an toto cells. cells. It Itis ispossible, possible, though though not not proven, proven, that that viruses viruses may may have have byby Transition Transition Year Year classes. classes. associatedassociated with with their their use? use? endospore?endospore? Describe Describe briefly briefly how how an an endospore endospore forms. forms. facilitatedfacilitated the the evolution evolution of of multicellular multicellular organisms. organisms. 7. 7.Compare Compare and and contrast contrast interferons interferons and and antibodies antibodies under under the the NameName two two types types of of heterotrophic heterotrophic nutrition nutrition used used by by bacteria. bacteria. headings;headings; chemical chemical nature, nature, speed speed of of action, action, specificity specificity of ofaction, action, howhow they they interact. interact. (c)(c) DistinguishDistinguish clearly clearly between between antibodies antibodies and and antibiotics antibiotics by by writing writing BiographicalBiographical Notes Notes LearningLearning Outcomes Outcomes a anote note about about each. each. In In relation relation to to antibodies, antibodies, distinguish distinguish between between 8.8. Write Write a ashort short account account of of how how phages phages have have been been used used in in the the activeactive and and passive passive immunity. immunity. OnOn completion completion of of this this lesson, lesson, students students should should be be able able to: to: treatmenttreatment of of bacterial bacterial disease. disease. What What advantage advantage might might bacteriophage bacteriophage AlickAlick Isaacs Isaacs (1921- (1921- 1967) 1967) UsingUsing your your knowledge knowledge of of antibiotics antibiotics and and bacteria, bacteria, suggest suggest why why therapytherapy have have over over antibiotic antibiotic treatment? treatment? HisHis paternal paternal grandparents grandparents migrated migrated to to England England from from Lithuania Lithuania about about a aperson person is ismore more likely likely to to pick pick up up an an infection infection in in hospital hospital than than • • DescribeDescribe how how modern modern biotechnology biotechnology allows allows the the efficient efficient massmass 1880.1880. The The family family moved moved to to Scotland Scotland where where his his father father Louis Louis opened opened a a productionproduction of of useful useful biochemicals biochemicals 9.9. Write Write a astory story as as told told by by an an interferon interferon molecule molecule beginning beginning with with the the atat home. home. shopshop in in Kilmarnock. Kilmarnock. Alick Alick graduated graduated in in medicine medicine from from Glasgow Glasgow in in 1944. 1944. wordswords “Suddenly “Suddenly I wasI was there....” there....” • • ExplainExplain why why obtaining obtaining biochemicals biochemicals by by other other means means may may be be LeavingLeaving Certificate Certificate Biology Biology 2010 2010 (HL) (HL) Q. Q.10 10 a, a,b b HisHis worked worked in in bacteriology bacteriology and and subsequently subsequently spent spent time time in in Australia Australia studyingstudying the the influenza influenza virus. virus. Research Research into into potential potential anti-viral anti-viral agents agents inefficient,inefficient, expensive expensive and and not not yield yield them them in in sufficient sufficient quantity quantity 10.10. Write Write a ashort short essay essay “A “A future future – –gene gene therapy therapy and and interferon” interferon” (a)(a) NameName the the base base in in DNA DNA that that pairs pairs with with cytosine. cytosine. waswas a agrowing growing field field and and he, he, with with Jean Jean Lindenmann, Lindenmann, isolated isolated interferon interferon • • OutlineOutline the the role role of of interferon interferon atat a aLondon London laboratory laboratory in in 1957. 1957. He He was was awarded awarded the the Bellahouston Bellahouston gold gold • • OutlineOutline some some medical medical uses uses of of interferons interferons WhatWhat are are the the two two main main events events in in the the replication replication of of DNA? DNA? medalmedal for for his his work work on on influenza influenza and and was was mademade a aFellow Fellow of ofthe the Royal Royal • • OutlineOutline the the process process involved involved in in using using micro-organisms micro-organisms to to produce produce (b)(b) ExplainExplain the the terms terms transcription transcription and and translation. translation. In In which which structures structures SocietySociety in in 1966. 1966. complexcomplex bio-molecules. bio-molecules. True/FalseTrue/False Questions Questions inin the the cell cell does does translation translation occur? occur? JeanJean Lindenmann Lindenmann HowHow many many bases bases in in sequence sequence make make up up a acodon codon in in mRNA? mRNA? Each Each a)a) InterferonInterferon is isproduced produced during during the the clotting clotting process process at at the the site site mRNAmRNA codon codon specifies specifies one one of ofthree three possible possible outcomes outcomes during during BornBorn Zagreb Zagreb 1924 1924 to to Swiss Swiss parents, parents, he he obtained obtained an an MD MD in in Zurich Zurich ofof a awound. wound. T T F F proteinprotein synthesis. synthesis. Name Name these these three three possible possible outcomes. outcomes. UniversityUniversity 1951. 1951. Working Working in in London London 1956-57 1956-57 he he isolated isolated interferon, interferon, GeneralGeneral Learning Learning Points Points notingnoting that that after after initial initial exposure exposure to to virus, virus, it ittook took time time for for tissues tissues to to b)b) TransferTransfer of of genetic genetic material material between between organisms organisms does does not not WhatWhat does does the the letter letter ‘t’ ‘t’ stand stand for for in in tRNA? tRNA? During During translation translation one one produceproduce it. it. Back Back in in Zurich, Zurich, he he later later showed showed that that a astrain strain of of mice mice immune immune TheseThese are are additional additional relevant relevant points points which which are are used used to to extend extend happenhappen in in nature. nature. T T F F endend of of a atRNA tRNA molecule molecule attaches attaches to to an an mRNA mRNA toto influenza influenza carried carried a adominant dominant autosomal autosomal gene gene Mx.Mx. He He further further knowledgeknowledge and and facilitate facilitate discussion. discussion. discovereddiscovered that that this this gene gene produced produced a aprotein protein that that stopped stopped viral viral c)c) BacteriaBacteria never never revert revert to to their their original original state state after after genetic genetic LeavingLeaving Certificate Certificate Biology Biology 2013 2013 (OL) (OL) Q. Q.10 10 b b replicationreplication and and that that the the gene gene action action was was activated activated by by interferon. interferon. • • InterferonsInterferons are are proteins proteins produced produced by by animals, animals, production production happening happening modification.modification. T T F F (b)(b) ThereThere are are four four bases bases in in DNA DNA onlyonly in in the the presence presence of of certain certain stimuli. stimuli. They They have have anti-viral anti-viral and and d)d) TheThe majority majority of of eukaryotic eukaryotic genes, genes, in in contrast contrast to to those those found found in in structure.structure. These These are are adenine adenine anti-canceranti-cancer properties. properties. Scientists Scientists have have classified classified them them based based on on the the prokaryotes,prokaryotes, have have introns. introns. T T F F (A),(A), cytosine cytosine (C), (C), thymine thymine (T) (T) differentdifferent interferon interferon receptors receptors found found on on cells. cells. e)e) E.coliE.coli are are the the only only bacteria bacteria that that can can be be successfully successfully modified modified andand guanine guanine (G). (G). Name Name the the • • TypeType 1 1includes includes IFN-α, IFN-α, β βand and other other variants. variants. toto produce produce interferon. interferon. T T F F basesbases at at positions positions X Xand and Y Y ReviseRevise The The Terms Terms • • TypeType 2. 2. Human Human IFN-γ IFN-γ belongs belongs to to this this group. group. Type Type 2 2is isproduced produced only only inin the the diagram. diagram. f) f) InIn a afirst-time first-time viral viral infection, infection, interferon interferon is isproduced produced moremore CanCan you you recall recall the the meaning meaning of of the the following following terms? terms? byby some some immune immune cells; cells; the the others others are are produced produced by by all all cells. cells. quicklyquickly than than antibodies. antibodies. T T F F WhereWhere in in a ahuman human cell cell would would RevisingRevising terminology terminology is is a apowerful powerful aid aid to to recall recall and and retention. retention. • • TypeType 3. 3. Most Most recently recently discovered; discovered; these these interferons interferons have have anti-viral anti-viral youyou expect expect to to find find mostmost g)g) CytokinesCytokines are are secreted secreted low-molecular low-molecular weight weight proteins proteins that that angiogenesis,angiogenesis, antibodies, antibodies, antigen, antigen, apoptosis, apoptosis, bacteria, bacteria, chromatography, chromatography, properties.properties. DNA?DNA? regulateregulate the the immune immune response. response. T T F F clone,clone, cloning cloning vectors, vectors, codons, codons, cytokines, cytokines, DNA, DNA, eukaryotic, eukaryotic, gene, gene, • • It Itis isnot not practical practical to to obtain obtain useful useful interferon interferon in in commercial commercial quantities quantities ProteinsProteins are are made made in in the the geneticgenetic engineering, engineering, genome, genome, glycoprotein, glycoprotein, glycosylation, glycosylation, hybrid hybrid fromfrom animals. animals. Interferons Interferons from from one one animal animal generally generally have have very very h)h) InterferonsInterferons were were being being used used clinically clinically on on a alarge large scale scale within within ribosomesribosomes using using a acode code from from DNA. DNA. Name Name the the molecule molecule that that interferons,interferons, inhibitors, inhibitors, interferon, interferon, introns, introns, lysogenic, lysogenic, lytic, lytic, MHC MHC antigens, antigens, limitedlimited effectiveness effectiveness in in others. others. Moreover, Moreover, natural natural levels levels in in tissues tissues tenten years years of of their their discovery discovery in in 1957. 1957. T T F F carriescarries the the DNA DNA code code to to the the ribosomes. ribosomes. monoclonalmonoclonal antibodies, antibodies, phages, phages, plasmids, plasmids, professional professional antigen- antigen- areare very very low. low. DNA DNA coding coding for for human human interferon interferon has has been been isolated, isolated, i) i) InflammationInflammation is isan an immune immune response response to to harmful harmful stimuli, stimuli, not not presentingpresenting cells, cells, prokaryotic, prokaryotic, receptors, receptors, recombinant recombinant DNA DNA technology, technology, purifiedpurified and and inserted inserted into into bacteria bacteria which which are are then then cloned cloned to to produce produce WhatWhat is ismeant meant by by DNA DNA profiling? profiling? In In DNA DNA profiling, profiling, what what is is used used alwaysalways infections, infections, which which can can be be modulated modulated by by interferons. interferons. T T F F RNA,RNA, tumour, tumour, viruses viruses interferoninterferon in in industrial industrial quantities. quantities. It Itis isused used as as an an injectable injectable drug drug to to toto cut cut the the DNA DNA strands strands into into fragments? fragments? Give Give two two applications applications of of CheckCheck your your answers answers to to these these questions questions on on www.sta.ie. www.sta.ie. treattreat a avariety variety of of illnesses. illnesses. DNADNA profiling. profiling. CheckCheck the the Glossary Glossary of of terms terms for for this this lesson lesson on on www.sta.ie www.sta.ie UsingUsing bacteria bacteria to to makemake a a biomolecule biomolecule Using bacteria to make a biomolecule

SyllabusSyllabus references references StudentStudent Activities Activities ExaminationExamination Questions Questions DidDid You You Know? Know?

TheThe main main syllabus syllabus references references for for the the lesson lesson are: are: 1. 1.Make Make a lista list of ofthe the different different human human interferons. interferons. LeavingLeaving Certificate Certificate Biology Biology 2013 2013 (HL) (HL) Q. Q.12 12 • • Interferon-γInterferon-γ has has been been used used effectively effectively in inthe the treatment treatment of of osteopetrosis,osteopetrosis, an an inherited inherited disorder disorder characterised characterised by by an an overgrowth overgrowth JuniorJunior Certificate Certificate Science Science 2. 2.Draw Draw a postera poster picture picture of ofan an interferon interferon molecule. molecule. (a)(a) A Avirus virus has has been been described described as as a piecea piece of ofgenetic genetic material material that that has has of ofbone bone which which leads leads to tothe the development development of ofdeafness deafness and and blindness. blindness. 1C8.1C8. MicrobiologyMicrobiology and and biotechnology biotechnology escapedescaped from from a cell.a cell. Give Give one one piece piece of ofevidence evidence that that supports supports this this ThereThere is isreduced reduced space space for for bone bone marrow marrow resulting resulting in inanaemia anaemia and and 3. 3.Draw Draw a picturea picture narrative narrative illustrating illustrating the the manufacture manufacture of ofinterferon, interferon, description.description. impairedimpaired resistance resistance to todisease. disease. fromfrom the the isolation isolation of ofthe the required required DNA DNA to toputting putting interferon interferon powder powder LeavingLeaving Certificate Certificate Biology Biology intointo a bottle.a bottle. VirusesViruses are are examples examples of ofobligate obligate parasites. parasites. Explain Explain why why this this is isthe the • • MultipleMultiple sclerosis sclerosis is isan an auto-immune auto-immune disorder disorder where where the the body’s body’s defencesdefences attack attack the the myelin myelin sheath sheath that that encloses encloses neurons. neurons. Impaired Impaired 2.1.4.2.1.4. ProkaryoticProkaryotic and and eukaryotic eukaryotic cells cells case.case. Give Give an an example example of ofhow how a virusa virus might might be be beneficial beneficial to to 4. 4.Why Why might might it beit be a gooda good idea idea to togive give interferon interferon by by injection injection rather rather mankind.mankind. nervenerve impulse impulse transmission transmission follows, follows, leading leading to toweakness weakness and and 2.52.5 GeneticsGenetics thanthan orally? orally? paralysis.paralysis. Interferon Interferon therapy therapy appears appears to towork work partly partly by by slowing slowing 3.1.1.3.1.1. DiversityDiversity of oforganisms organisms (b)(b) NameName the the kingdom kingdom to towhich which bacteria bacteria belong. belong. Draw Draw a largea large diagram diagram downdown the the production production of ofT-cells T-cells 5.5. E.coli E.coli bacteria bacteria can can make make interferon. interferon. So So can can yeast yeast cells. cells. There There is is of ofa bacteriala bacterial cell cell to toshow: show: • • DiphtheriaDiphtheria and and botulism botulism toxins toxins are are produced produced by by genes genes inserted inserted by by 3.1.2.3.1.2. Micro-organismsMicro-organisms alsoalso experimentation experimentation with with cell-free cell-free methods. methods. Can Can you you think think of of 1. 1.The The relative relative positions positions of ofthe the cell cell wall, wall, cell cell membrane membrane and and virusesviruses into into bacteria. bacteria. Scientists Scientists have have also also discovered discovered that that genes genes 3.1.3.3.1.3. Monera,Monera, e.g. e.g. bacteria bacteria advantages/disadvantagesadvantages/disadvantages associated associated with with each each method? method? capsule.capsule. 2. 2.A Aplasmid. plasmid. veryvery similar similar to tothose those coding coding for for some some fusion fusion proteins proteins in inhuman human cells cells 3.1.9.3.1.9. NatureNature of ofbacteria bacteria and and fungi. fungi. areare also also found found in inviruses. viruses. These These proteins proteins enable enable cells cells to toorganise organise 6. 6.Plasmids Plasmids and and phages phages are are vectors vectors for for introducing introducing DNA DNA into into bacterial, bacterial, LabelLabel each each of ofthe the above above structures. structures. andand other, other, cells. cells. The The future future may may see see increased increased use use of ofcosmids cosmids and and intointo tissues tissues and and organs organs whereas whereas the the virus virus uses uses them them to togain gain access access ScienceScience and and Technology Technology in in Action Action is isalso also widely widely used used artificialartificial chromosomes. chromosomes. WhatWhat are are the the drawbacks drawbacks and and prospects prospects UnderUnder what what circumstances circumstances does does a bacteriala bacterial cell cell form form an an to tocells. cells. It isIt ispossible, possible, though though not not proven, proven, that that viruses viruses may may have have byby Transition Transition Year Year classes. classes. associatedassociated with with their their use? use? endospore?endospore? Describe Describe briefly briefly how how an anendospore endospore forms. forms. facilitatedfacilitated the the evolution evolution of ofmulticellular multicellular organisms. organisms. 7. 7.Compare Compare and and contrast contrast interferons interferons and and antibodies antibodies under under the the NameName two two types types of ofheterotrophic heterotrophic nutrition nutrition used used by by bacteria. bacteria. headings;headings; chemical chemical nature, nature, speed speed of ofaction, action, specificity specificity of ofaction, action, howhow they they interact. interact. (c)(c) DistinguishDistinguish clearly clearly between between antibodies antibodies and and antibiotics antibiotics by by writing writing BiographicalBiographical Notes Notes LearningLearning Outcomes Outcomes a notea note about about each. each. In Inrelation relation to toantibodies, antibodies, distinguish distinguish between between 8. 8.Write Write a shorta short account account of ofhow how phages phages have have been been used used in inthe the activeactive and and passive passive immunity. immunity. OnOn completion completion of ofthis this lesson, lesson, students students should should be be able able to: to: treatmenttreatment of ofbacterial bacterial disease. disease. What What advantage advantage might might bacteriophage bacteriophage AlickAlick Isaacs Isaacs (1921- (1921- 1967) 1967) UsingUsing your your knowledge knowledge of ofantibiotics antibiotics and and bacteria, bacteria, suggest suggest why why therapytherapy have have over over antibiotic antibiotic treatment? treatment? HisHis paternal paternal grandparents grandparents migrated migrated to toEngland England from from Lithuania Lithuania about about a persona person is ismore more likely likely to topick pick up up an an infection infection in inhospital hospital than than • • DescribeDescribe how how modern modern biotechnology biotechnology allows allows the the efficient efficient mass mass 1880.1880. The The family family moved moved to toScotland Scotland where where his his father father Louis Louis opened opened a a productionproduction of ofuseful useful biochemicals biochemicals 9. 9.Write Write a storya story as as told told by by an an interferon interferon molecule molecule beginning beginning with with the the at athome. home. shopshop in inKilmarnock. Kilmarnock. Alick Alick graduated graduated in inmedicine medicine from from Glasgow Glasgow in in1944. 1944. wordswords “Suddenly “Suddenly I was I was there....” there....” • • ExplainExplain why why obtaining obtaining biochemicals biochemicals by by other other means means may may be be LeavingLeaving Certificate Certificate Biology Biology 2010 2010 (HL) (HL) Q. Q.10 10a, a,b b HisHis worked worked in inbacteriology bacteriology and and subsequently subsequently spent spent time time in inAustralia Australia studyingstudying the the influenza influenza virus. virus. Research Research into into potential potential anti-viral anti-viral agents agents inefficient,inefficient, expensive expensive and and not not yield yield them them in insufficient sufficient quantity quantity 10.10. Write Write a shorta short essay essay “A “A future future – gene– gene therapy therapy and and interferon” interferon” (a)(a) NameName the the base base in inDNA DNA that that pairs pairs with with cytosine. cytosine. waswas a growinga growing field field and and he, he, with with Jean Jean Lindenmann, Lindenmann, isolated isolated interferon interferon • • OutlineOutline the the role role of ofinterferon interferon at ata Londona London laboratory laboratory in in1957. 1957. He He was was awarded awarded the the Bellahouston Bellahouston gold gold • • OutlineOutline some some medical medical uses uses of ofinterferons interferons WhatWhat are are the the two two main main events events in inthe the replication replication of ofDNA? DNA? medalmedal for for his his work work on on influenza influenza and and was was made made a aFellow Fellow of theof the Royal Royal • • OutlineOutline the the process process involved involved in inusing using micro-organisms micro-organisms to toproduce produce (b)(b) ExplainExplain the the terms terms transcription transcription and and translation. translation. In Inwhich which structures structures SocietySociety in in1966. 1966. complexcomplex bio-molecules. bio-molecules. True/FalseTrue/False Questions Questions in inthe the cell cell does does translation translation occur? occur? JeanJean Lindenmann Lindenmann HowHow many many bases bases in insequence sequence make make up up a codona codon in inmRNA? mRNA? Each Each a) a) InterferonInterferon is isproduced produced during during the the clotting clotting process process at atthe the site site mRNAmRNA codon codon specifies specifies one one of threeof three possible possible outcomes outcomes during during BornBorn Zagreb Zagreb 1924 1924 to toSwiss Swiss parents, parents, he he obtained obtained an an MD MD in inZurich Zurich of ofa wound.a wound. T T F F proteinprotein synthesis. synthesis. Name Name these these three three possible possible outcomes. outcomes. UniversityUniversity 1951. 1951. Working Working in inLondon London 1956-57 1956-57 he he isolated isolated interferon, interferon, GeneralGeneral Learning Learning Points Points notingnoting that that after after initial initial exposure exposure to tovirus, virus, it tookit took time time for for tissues tissues to to b) b) TransferTransfer of ofgenetic genetic material material between between organisms organisms does does not not WhatWhat does does the the letter letter ‘t’ ‘t’stand stand for for in intRNA? tRNA? During During translation translation one one produceproduce it. it.Back Back in inZurich, Zurich, he he later later showed showed that that a straina strain of ofmice mice immune immune TheseThese are are additional additional relevant relevant points points which which are are used used to toextend extend happenhappen in innature. nature. T T F F endend of ofa tRNAa tRNA molecule molecule attaches attaches to toan an mRNA mRNA to toinfluenza influenza carried carried a dominanta dominant autosomal autosomal gene gene Mx. Mx. He Hefurther further knowledgeknowledge and and facilitate facilitate discussion. discussion. discovereddiscovered that that this this gene gene produced produced a proteina protein that that stopped stopped viral viral c) c) BacteriaBacteria never never revert revert to totheir their original original state state after after genetic genetic LeavingLeaving Certificate Certificate Biology Biology 2013 2013 (OL) (OL) Q. Q.10 10b b replicationreplication and and that that the the gene gene action action was was activated activated by by interferon. interferon. • • InterferonsInterferons are are proteins proteins produced produced by by animals, animals, production production happening happening modification.modification. T T F F (b)(b) ThereThere are are four four bases bases in inDNA DNA onlyonly in inthe the presence presence of ofcertain certain stimuli. stimuli. They They have have anti-viral anti-viral and and d)d) TheThe majority majority of ofeukaryotic eukaryotic genes, genes, in incontrast contrast to tothose those found found in in structure.structure. These These are are adenine adenine anti-canceranti-cancer properties. properties. Scientists Scientists have have classified classified them them based based on onthe the prokaryotes,prokaryotes, have have introns. introns. T T F F (A),(A), cytosine cytosine (C), (C), thymine thymine (T) (T) differentdifferent interferon interferon receptors receptors found found on on cells. cells. e)e) E.coliE.coli are are the the only only bacteria bacteria that that can can be be successfully successfully modified modified andand guanine guanine (G). (G). Name Name the the • • TypeType 1 includes1 includes IFN-α, IFN-α, β andβ and other other variants. variants. to toproduce produce interferon. interferon. T T F F basesbases at atpositions positions X Xand and Y Y ReviseRevise The The Terms Terms • • TypeType 2. 2.Human Human IFN-γ IFN-γ belongs belongs to tothis this group. group. Type Type 2 is2 isproduced produced only only in inthe the diagram. diagram. f) f) In Ina first-timea first-time viral viral infection, infection, interferon interferon is isproduced produced more more CanCan you you recall recall the the meaning meaning of of the the following following terms? terms? byby some some immune immune cells; cells; the the others others are are produced produced by by all all cells. cells. quicklyquickly than than antibodies. antibodies. T T F F WhereWhere in ina humana human cell cell would would RevisingRevising terminology terminology is isa powerfula powerful aid aid to to recall recall and and retention. retention. • • TypeType 3. 3.Most Most recently recently discovered; discovered; these these interferons interferons have have anti-viral anti-viral youyou expect expect to tofind find most most g) g) CytokinesCytokines are are secreted secreted low-molecular low-molecular weight weight proteins proteins that that angiogenesis,angiogenesis, antibodies, antibodies, antigen, antigen, apoptosis, apoptosis, bacteria, bacteria, chromatography, chromatography, properties.properties. DNA?DNA? regulateregulate the the immune immune response. response. T T F F clone,clone, cloning cloning vectors, vectors, codons, codons, cytokines, cytokines, DNA, DNA, eukaryotic, eukaryotic, gene, gene, • • It isIt isnot not practical practical to toobtain obtain useful useful interferon interferon in incommercial commercial quantities quantities ProteinsProteins are are made made in inthe the geneticgenetic engineering, engineering, genome, genome, glycoprotein, glycoprotein, glycosylation, glycosylation, hybrid hybrid fromfrom animals. animals. Interferons Interferons from from one one animal animal generally generally have have very very h) h) InterferonsInterferons were were being being used used clinically clinically on on a largea large scale scale within within ribosomesribosomes using using a codea code from from DNA. DNA. Name Name the the molecule molecule that that interferons,interferons, inhibitors, inhibitors, interferon, interferon, introns, introns, lysogenic, lysogenic, lytic, lytic, MHC MHC antigens, antigens, limitedlimited effectiveness effectiveness in inothers. others. Moreover, Moreover, natural natural levels levels in intissues tissues tenten years years of oftheir their discovery discovery in in1957. 1957. T T F F carriescarries the the DNA DNA code code to tothe the ribosomes. ribosomes. monoclonalmonoclonal antibodies, antibodies, phages, phages, plasmids, plasmids, professional professional antigen- antigen- areare very very low. low. DNA DNA coding coding for for human human interferon interferon has has been been isolated, isolated, i) i) InflammationInflammation is isan animmune immune response response to toharmful harmful stimuli, stimuli, not not presentingpresenting cells, cells, prokaryotic, prokaryotic, receptors, receptors, recombinant recombinant DNA DNA technology, technology, purifiedpurified and and inserted inserted into into bacteria bacteria which which are are then then cloned cloned to toproduce produce WhatWhat is ismeant meant by by DNA DNA profiling? profiling? In InDNA DNA profiling, profiling, what what is isused used alwaysalways infections, infections, which which can can be be modulated modulated by by interferons. interferons. T T F F RNA,RNA, tumour, tumour, viruses viruses interferoninterferon in inindustrial industrial quantities. quantities. It isIt isused used as as an an injectable injectable drug drug to to to tocut cut the the DNA DNA strands strands into into fragments? fragments? Give Give two two applications applications of of CheckCheck your your answers answers to to these these questions questions on on www.sta.ie. www.sta.ie. treattreat a varietya variety of ofillnesses. illnesses. DNADNA profiling. profiling. CheckCheck the the Glossary Glossary of of terms terms for for this this lesson lesson on on www.sta.ie www.sta.ie