DOCSLIB.ORG

What Is Molecular Biology?

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
What Is Molecular Biology? Common Molecular Biology Techniques and Their Uses in Chemistry Becky Farmer Long Literature Seminar August 25, 2008 What is Molecular Biology? •Molecular biology is commonly defined as the branch of biology that examines biological phenomena at the molecular level •Mainly concerned with the study of biological macromolecules, such as DNA, RNA and proteins and their various interactions •Term “molecular biology” was coined in 1938 by Warren Weaver, the former director of the Division of Natural Sciences at the Rockefeller Foundation •Characterized by several common techniques: •Polymerase chain reaction (PCR) •Polyacrylamide gel electophoresis and blotting •Fluorescence activated cell sorting (FACS) •Knockout mice •RNA interference (RNAi) •Microarrays •Confocal microscopy •Fluorescent in situ hybridization (FISH) •Expression cloning/cell transfection •And many others….. The Basics: DNA, RNA and Everything In Between •The process by which genetic information is shared and expressed in the cell is known as the “central dogma of molecular biology” •DNA RNA protein •DNA is duplicated through the process of replication •mRNA is made through transcription •The genetic code is read from the mRNA by ribosomes, which synthesize proteins through the process of translation The Basics: DNA Replication •Genetic information is passed on to new cells via the process of semi-conservative replication •Each DNA strand containing the genetic code is copied by DNA polymerase III in a 5’3’ fashion .The leading strand is copied continuously, while the lagging strand is copied in short pieces (Okazaki fragments) and then ligated together •DNA helicase unwinds the leading edge of the replication fork, while topoisomerase prevents tangling of the chromosomal material Polymerase Chain Reaction (PCR) •Developed in 1983 by Kary Mullis, who received a Ph.D. in biochemistry from the University of California at Berkeley •Mullis conceived of the idea for PCR while working for Cetus Corporation, a biotechnology company located in Emeryville, CA •Mullis received the Nobel Prize in Chemistry for his work in 1993, the first prize ever awarded for research performed at a biotechnology company •In the press release for the Nobel Prize, the Royal Swedish Academy of Sciences described the impact of PCR as follows: Kary Mullis The biomedical applications of the PCR method are already legion. Now that it is possible to discover very small amounts of foreign DNA in an organism, viral and bacterial infections can be diagnosed without the time-consuming culture of microorganisms from patient samples. PCR is now being used, for example, to discover HIV infections. The method can also be exploited to localize the genetic alterations underlying hereditary diseases. Thus PCR, like site-directed mutagenesis, has a great potential within gene therapy. […] In police investigations PCR can give decisive information since it is now possible to analyze the DNA in a single drop of blood or in a hair found at the scene of a crime. Mechanism of PCR •PCR is used to exponentially amplify a sequence of DNA, known as the target sequence •Isolated DNA is added to reaction along with primers (complementary to flanking regions of target sequence), dNTPs, and heat-stable DNA Taq polymerase •First step involves heating the reaction to ~95 °C for 20-30 seconds to denature the double helix •Rapid cooling to 50-65 °C allows for annealing of primers •Reaction is then heated to 72 °C, the optimal temperate for the Taq polymerase, which then synthesizes new DNA strands from primers in 5’3’ direction Mechanism of PCR Continued •Another cycle then proceeds with the same temperature changes, or “steps”, with the newly synthesized DNA strands acting as templates •After the second cycle, short strands containing only the DNA sequence of interest are formed •These short strands can act as templates for further cycles, thereby leading to exponential amplification of the sequence of interest •Consists of generally 20-40 cycles, with each cycle consisting of discrete temperature steps •Ideal situation is amplification of 2n with n equal to the number of cycles .Amplification is a millionfold after 20 cycles and a billionfold after 30 cycles Quantitative Real-Time PCR (qRT-PCR) •Quantitation of gene expression levels is important for understanding of cell biology •Use of fluorescence emission generated during each cycle allows accurate quantitation of gene of interest •Most commonly used method involves fluorescent probe that hybridizes to sequence of interest •When Taq polymerase reaches probe, 5’ 3’ exonuclease activity destroys the probe and releases fluorescent dye Mocellin, S.; Rossi, C. R.; Pilati, P.; Nitti, D.; Marincola, F. M. Trends Mol. Med. 2003, 9, 189-195. The Basics: Transcription •Transcription is the process by which the genetic code in DNA is passed on to single- stranded mRNA .Particularly important step in gene expression since this is the level at which the cell regulates which proteins are made •RNA polymerase (I, II or III) binds to a promoter in the DNA sequence with the assistance of multiple transcription factors and synthesizes the gene .Promoter usually 25 nucleotides upstream of the gene of interest •DNA helix reassembles after transcription, while mRNA is extruded from the nucleus, post-transcriptionally modified and translated into protein Reverse Transcription PCR (RT-PCR) •Reverse transcription is the process used by viruses to synthesize DNA from RNA genome •Similar principle is used for reverse transcription PCR to convert RNA to cDNA •Technique used for measurement of gene expression in the cell •Often combined with quantitative real-time PCR for accurate picture of actions occurring inside cell Bustin, S. A.; Mueller, R. Clin. Sci. 2005, 109, 365-379. Electrochemical Real-Time PCR •Current real-time PCR protocols rely on bulky optical systems for detection of amplification •For point-of-care applications, need an inexpensive portable device •In the first step, PCR is carried out in solution phase to give dsDNA containing modified Fc markers •The DNA is then annealed to an electrode containing complementary probes •Extension occurs, increasing number of Fc markers on surface electrical signal Yeung, S. S. W.; Lee, T. M. H.; Hsing, I. J. Am. Chem. Soc. 2006, 128, 13374-13375. Electrochemical Real-Time PCR Continued •Voltammetric scans taken in presence (sample) and absence (negative control) of target sequence demonstrate the buildup of Fc markers on glass electrode •Current measured as a function of cycle number shows that signal is detected on a real-time basis •Signal is detected more quickly than in traditional optical systems could be used for detection of small amounts of DNA Yeung, S. S. W.; Lee, T. M. H.; Hsing, I. J. Am. Chem. Soc. 2006, 128, 13374-13375. Polyacrylamide Gel Electrophoresis (PAGE) •Gel electrophoresis is used to separate DNA, RNA or proteins by size and charge •The gel is made of a crosslinked polyacrylamide, although it can also be made of agarose •Current is applied to the gel, which causes the proteins/nucleic acids of interest to migrate toward anode/cathode causes discrete bands which can be identified by size •SDS-PAGE is used to analyze proteins, since sodium dodecyl sulfate denatures proteins Blotting •Gel electrophoresis is the first step in the biological procedure known as blotting •Once macromolecules of interest have been separated using gel electrophoresis, they are transferred to a separate surface where they are further analyzed Edwin Southern •Types of blots: .Southern blot – used to probe for a specific sequence of DNA, named after Edwin Southern, a British biologist who developed the technique in 1975 .Western blot – analysis of proteins via antibody detection .Northern blot – detection of specific sequences of RNA The Basics: Translation •Translation is the process by which the genetic code found in mRNA is used to synthesize proteins •Codons (sequence of three nucleotides) are matched to their corresponding tRNA carrying the appropriate amino acid •The ribosome catalyzes the elongation of the growing peptide chain and then transports the nascent protein into the endoplasmic reticulum, where it is folded into its 3D structure and post-translationally modified Western Blots •Western blots are the most commonly used blotting technique in molecular biology •Used to gauge the level of protein expression in the cell since protein levels are a result of gene expression, Western blot gives accurate depiction of gene activity in the cell •First developed by George Stark at Stanford University, then later perfected and termed “western blot” by W. Neal Burnette in 1981 •Western blotting is used to diagnose several diseases, including HIV and bovine spongiform encephalopathy Western Blots in Hsp90 Inhibitor Evaluation •Hsp90 is one of the most abundant proteins in humans accounts for 1-2% of all protein •Assists in protein folding as a chaperone and is also important in tumor progression .Stabilizes mutant oncogenic proteins such as p53 and bcr/abl .Maintains PI3K and AKT, both of which inhibit apoptosis .Promotes metastasis (MMP-2) and angiogenesis (NOS and VEGF) Yu, X. M.; Shen, G.; Neckers, L.; Blake, H.; Holzbeierlein, J.; Cronk, B.; Blagg, B. S. J. J. Am. Chem. Soc. 2005, 127, 12778-12779. Hsp90 Inhibitor Development Continued •Each compound was tested for its ability to cause degradation of Hsp90 client protein phospho-AKT in SKBR3 breast cancer cells •Geldanamycin was used as positive control •SAR shows several patterns: .7-position of coumarin ring is important for activity (B vs. D/E) .Compounds with amide linker show more potent action (A) .Diol may mimic ATP in binding site (4) •Compound A4 was identified as the most potent inhibitor Yu, X. M.; Shen, G.; Neckers, L.; Blake, H.; Holzbeierlein, J.; Cronk, B.; Blagg, B. S. J. J. Am. Chem. Soc. 2005, 127, 12778-12779. Hsp90 Inhibitor Development Continued •Novobiocin analog A4 also tested to determine their action against hormone receptors •Two cell lines were used: LNCap (mutated androgen receptor PCa) and LAPC-4 (wild-type androgen receptor PCa) •A4 causes degradation of androgen receptor (AR), AKT and HIF-1α at low concentrations (~1µM) Yu, X.
Load more

Recommended publications
  • USA Education Ph.D., Biology, Massachusetts Institute of Tech
    Victor R. Ambros, Ph.D. Silverman Professor of Natural Sciences Program in Molecular Medicine University of Massachusetts Medical School373 Plantation Street, Suite 306 Worcester, MA 01605 (508) 856-6380 [email protected] Personal Born: Hanover, NH, USA on December 1, 1953 Citizenship: USA Education Ph.D., Biology, Massachusetts Institute of Technology, Cambridge, MA 1976-1979 Thesis Title: The protein covalently linked to the 5' end of poliovirus RNA Advisor: Dr. David Baltimore B.S., Biology, Massachusetts Institute of Technology, Cambridge, MA 1971-1975 Professional Appointments Silverman Professor of Natural Sciences 2009-present Co-Director, RNA Therapeutics Institute 2009-2016 Professor, Program in Molecular Medicine 2008-present University of Massachusetts Medical School, Worcester, MA Professor of Genetics, Dartmouth Medical School 2001-2007 Professor, Biological Sciences, Dartmouth Medical School 1996-2001 Associate Professor, Biological Sciences, Dartmouth Medical School 1992-1996 Associate Professor, Department of Cellular and Development Biology, 1988-1992 Harvard University, Cambridge, MA Assistant Professor, Department of Cellular and Development Biology, 1985-1988 Harvard University, Cambridge, MA Postdoctoral Research 1979-1985 Supervisor: Dr. H. Robert Horvitz Massachusetts Institute of Technology, Cambridge, MA Graduate Research 1976-1979 Supervisor: Dr. David Baltimore Massachusetts Institute of Technology, Cambridge, MA Research Assistant 1975-1976 Supervisor: Dr. David Baltimore Center for Cancer Research,
    [Show full text]
  • Biochemistry and the Genomic Revolution 1.1
    Dedication About the authors Preface Tools and Techniques Clinical Applications Molecular Evolution Supplements Supporting Biochemistry, Fifth Edition Acknowledgments I. The Molecular Design of Life 1. Prelude: Biochemistry and the Genomic Revolution 1.1. DNA Illustrates the Relation between Form and Function 1.2. Biochemical Unity Underlies Biological Diversity 1.3. Chemical Bonds in Biochemistry 1.4. Biochemistry and Human Biology Appendix: Depicting Molecular Structures 2. Biochemical Evolution 2.1. Key Organic Molecules Are Used by Living Systems 2.2. Evolution Requires Reproduction, Variation, and Selective Pressure 2.3. Energy Transformations Are Necessary to Sustain Living Systems 2.4. Cells Can Respond to Changes in Their Environments Summary Problems Selected Readings 3. Protein Structure and Function 3.1. Proteins Are Built from a Repertoire of 20 Amino Acids 3.2. Primary Structure: Amino Acids Are Linked by Peptide Bonds to Form Polypeptide Chains 3.3. Secondary Structure: Polypeptide Chains Can Fold Into Regular Structures Such as the Alpha Helix, the Beta Sheet, and Turns and Loops 3.4. Tertiary Structure: Water-Soluble Proteins Fold Into Compact Structures with Nonpolar Cores 3.5. Quaternary Structure: Polypeptide Chains Can Assemble Into Multisubunit Structures 3.6. The Amino Acid Sequence of a Protein Determines Its Three-Dimensional Structure Summary Appendix: Acid-Base Concepts Problems Selected Readings 4. Exploring Proteins 4.1. The Purification of Proteins Is an Essential First Step in Understanding Their Function 4.2. Amino Acid Sequences Can Be Determined by Automated Edman Degradation 4.3. Immunology Provides Important Techniques with Which to Investigate Proteins 4.4. Peptides Can Be Synthesized by Automated Solid-Phase Methods 4.5.
    [Show full text]
  • Nature Medicine Essay
    COMMENTARY LASKER BASIC MEDICAL RESEARCH AWARD Of maize and men, or peas and people: case histories to justify plants and other model systems David Baulcombe One of the byproducts of molecular biology cork is altogether filled with air, and that air is has been support for the ‘model system’ con- perfectly enclosed in little boxes or cells distinct cept. All living organisms are based on the same from one another.”)2 (Fig. 1). Two hundred fifty genetic code, they have similar subcellular years later, Beijerinck discovered a contagium structures and they use homologous metabolic vivum fluidum in extracts of diseased tobacco pathways. So, mechanisms can be investigated plants that he later referred to as a virus3. using organisms other than those in which In contemporary science, a green alga— the knowledge will be exploited for practical Chlamydomonas reinhardtii—is a useful model benefit. Model systems are particularly use- in the analysis of kidney disease4. However, ful in the early discovery phase of a scientific in this article, I refer to the contribution of endeavor, and recent progress in biomedical plant biology to a family of mechanisms that I science has fully vindicated their use. Jacques refer to as RNA silencing. This topic has been Monod, for example, famously justified his reviewed comprehensively elsewhere5,6, so here work on a bacterial model system by stating I focus on personal experience and my view of that “what is true for Escherichia coli is also future potential from this work. true for elephants.” My fellow laureates, Victor Ambros and Gary Ruvkun, can defend the use The early history of RNA silencing in of the worm Caenorhabditis elegans as a good plants model system and so I will focus on plants.
    [Show full text]
  • Blotting Techniques Blotting Is the Technique in Which Nucleic Acids Or
    Blotting techniques Blotting is the technique in which nucleic acids or proteins are immobilized onto a solid support generally nylon or nitrocellulose membranes. Blotting of nucleic acid is the central technique for hybridization studies. Nucleic acid labeling and hybridization on membranes have formed the basis for a range of experimental techniques involving understanding of gene expression, organization, etc. Identifying and measuring specific proteins in complex biological mixtures, such as blood, have long been important goals in scientific and diagnostic practice. More recently the identification of abnormal genes in genomic DNA has become increasingly important in clinical research and genetic counseling. Blotting techniques are used to identify unique proteins and nucleic acid sequences. They have been developed to be highly specific and sensitive and have become important tools in both molecular biology and clinical research. General principle The blotting methods are fairly simple and usually consist of four separate steps: electrophoretic separation of protein or of nucleic acid fragments in the sample; transfer to and immobilization on paper support; binding of analytical probe to target molecule on paper; and visualization of bound probe. Molecules in a sample are first separated by electrophoresis and then transferred on to an easily handled support medium or membrane. This immobilizes the protein or DNA fragments, provides a faithful replica of the original separation, and facilitates subsequent biochemical analysis. After being transferred to the support medium the immobilized protein or nucleic acid fragment is localized by the use of probes, such as antibodies or DNA, that specifically bind to the molecule of interest. Finally, the position of the probe that is bound to the immobilized target molecule is visualized usually by autoradiography.
    [Show full text]
  • Southern Blotting Teacher’S Guidebook
    PR139 G-Biosciences ♦ 1-800-628-7730 ♦ 1-314-991-6034 ♦ [email protected] A Geno Technology, Inc. (USA) brand name Southern Blotting Teacher’s Guidebook (Cat. # BE-315) think proteins! think G-Biosciences www.GBiosciences.com MATERIALS INCLUDED ....................................................................................................... 3 SPECIAL HANDLING INSTRUCTIONS ................................................................................... 3 ADDITIONAL EQUIPMENT REQUIRED ................................................................................ 3 TIME REQUIRED ................................................................................................................. 3 AIMS .................................................................................................................................. 4 BACKGROUND ................................................................................................................... 4 TEACHER’S PRE EXPERIMENT SET UP ................................................................................ 5 PREPARATION OF AGAROSE GEL ................................................................................... 5 PREPARE THE DNA LADDER ........................................................................................... 5 PREPARE DILUTE DNA STAINING SOLUTION .................................................................. 5 MATERIALS FOR DEMONSTRATION ................................................................................... 6 DEMONSTRATION
    [Show full text]
  • Ethical Principles in Ethical Principles in Scientific Research Scientific Research and Publications
    Hacettepe University Institute of Oncology Library ETHICAL PRINCIPLES IN SCIENTIFIC LectureRESEARCH author AND PUBLICATIONSOnlineby © Emin Kansu,M.D.,FACP ESCMID ekansu@ada. net. tr Library Lecture author Onlineby © ESCMID HACETTEPE UNIVERSITY MEDICAL CENTER Library Lecture author Onlineby © ESCMID INSTITUTE OF ONCOLOGY - HACETTEPE UNIVERSITY Ankara PRESENTATION • UNIVERSITY and RESEARCHLibrary • ETHICS – DEFINITION • RESEARCH ETHICS • PUBLICATIONLecture ETHICS • SCIENTIFIC MISCONDUCTauthor • SCIENTIFIC FRAUD AND TYPES Onlineby • HOW TO PREVENT© UNETHICAL ISSUES • WHAT TO DO FOR SCIENTIFIC ESCMIDMISCONDUCTS Library Lecture author Onlineby © ESCMID IMPACT OF TURKISH SCIENTISTS 85 % University – Based 1.78% 0.2 % 19. 300 18th ‘ACADEMIA’ UNIVERSITY Library AN INSTITUTION PRODUCING and DISSEMINATING SCIENCE Lecture BASIC FUNCTIONS author Online-- EDUCATIONby © -- RESEARCH ESCMID - SERVICESERVICE UNIVERSITY Library • HAS TO BE OBJECTIVE • HAS TO BE HONESTLecture AND ETHICAL • HAS TO PERFORM THEauthor “STATE-OF-THE ART” • HAS TO PLAYOnline THEby “ROLE MODEL” , TO BE GENUINE AND ©DISCOVER THE “NEW” • HAS TO COMMUNICATE FREELY AND HONESTLY WITH ALL THE PARTIES INVOLVED IN ESCMIDPUBLIC WHY WE DO RESEARCH ? Library PRIMARY AIM - ORIGINAL CONTRIBUTIONS TO SCIENCE Lecture SECONDARY AIM author - TOOnline PRODUCEby A PAPER - ACADEMIC© PROMOTION - TO OBTAIN AN OUTSIDE SUPPORT ESCMID SCIENTIFIC RESEARCH Library A Practice aimed to contribute to knowledge or theoryLecture , performed in disciplined methodologyauthor and Onlineby systematic approach©
    [Show full text]
  • PIE-1 Sumoylation Promotes Germline Fates and Pirna-Dependent Silencing in C
    RESEARCH ARTICLE PIE-1 SUMOylation promotes germline fates and piRNA-dependent silencing in C. elegans Heesun Kim1†, Yue-He Ding1†, Shan Lu2, Mei-Qing Zuo2, Wendy Tan1, Darryl Conte Jr1, Meng-Qiu Dong2, Craig C Mello1,3* 1RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States; 2National Institute of Biological Sciences, Beijing, China; 3Howard Hughes Medical Institute, Chevy Chase, United States Abstract Germlines shape and balance heredity, integrating and regulating information from both parental and foreign sources. Insights into how germlines handle information have come from the study of factors that specify or maintain the germline fate. In early Caenorhabditis elegans embryos, the CCCH zinc finger protein PIE-1 localizes to the germline where it prevents somatic differentiation programs. Here, we show that PIE-1 also functions in the meiotic ovary where it becomes SUMOylated and engages the small ubiquitin-like modifier (SUMO)-conjugating machinery. Using whole-SUMO-proteome mass spectrometry, we identify HDAC SUMOylation as a target of PIE-1. Our analyses of genetic interactions between pie-1 and SUMO pathway mutants suggest that PIE-1 engages the SUMO machinery both to preserve the germline fate in the embryo and to promote Argonaute-mediated surveillance in the adult germline. *For correspondence: Introduction [email protected] During every life cycle, the eukaryotic germline orchestrates a remarkable set of informational tasks †These authors contributed that shape heredity and create variation necessary for the evolution of new species. One approach equally to this work for understanding the mechanisms that promote germline specification and function has been the identification of genes whose protein products localize exclusively to the germline and for which Competing interests: The loss-of-function mutations result in absent or non-functional germ cells and gametes (Seydoux and authors declare that no Braun, 2006).
    [Show full text]
  • Bioanalytical Chemistry 4. Gel Electrophoresis
    73 Bioanalytical chemistry 4. Gel Electrophoresis Required reading: Sections 9.1, 9.2.3, 9.2.4, 9.5.1, 10.1 to 10.7, 11.1 to 11.5, and 15.5 of Mikkelsen and Cortón, Bioanalytical Chemistry Some objectives for this section ⇒ You will know what DNA agarose gel electrophoresis is ⇒ You will know what the difference between normal and pulsed field electrophoresis of DNA ⇒ You will know what SDS-PAGE is. ⇒ You will understand how the basis for molecular basis for size-based separation of proteins by SDS- PAGE. ⇒ You will know what IEF is. ⇒ You will know how SDS-PAGE and IEF can be combined in 2-dimensional gel electroophoresis ⇒ You will know what a Western Blot is. ⇒ You will appreciate how these techniques can be used in the analysis of DNA and proteins. Primary Source Material • Chapter 4 and 6 of Biochemistry: Berg, Jeremy M.; Tymoczko, John L.; and Stryer, Lubert (NCBI bookshelf). • Chapter 3 and 7 of Molecular Cell Biology 4th ed. (Ch. 9, 5th ed.): Lodish, Harvey; Berk, Arnold; Zipursky, S. Lawrence; Matsudaira, Paul; Baltimore, David; Darnell, James E. (NCBI bookshelf). • Chapter 12 of Introduction to Genetic Analysis Anthony: J.F. Griffiths, Jeffrey H. Miller, David T. Suzuki, Richard C. Lewontin, William M. Gelbart (NCBI bookshelf). • Some animations are from http://www.wiley-vch.de/books/info/3-527-30300-6/. • http://www.piercenet.com/ Electrophoresis 74 The velocity of migration (v) of a molecule in an electric field depends on the electric field strength (E), the net charge on the protein (z), and the frictional coefficient (f).
    [Show full text]
  • Fire Departments of Pathology and Genetics, Stanford University School of Medicine, 300 Pasteur Drive, Room L235, Stanford, CA 94305-5324, USA
    GENE SILENCING BY DOUBLE STRANDED RNA Nobel Lecture, December 8, 2006 by Andrew Z. Fire Departments of Pathology and Genetics, Stanford University School of Medicine, 300 Pasteur Drive, Room L235, Stanford, CA 94305-5324, USA. I would like to thank the Nobel Assembly of the Karolinska Institutet for the opportunity to describe some recent work on RNA-triggered gene silencing. First a few disclaimers, however. Telling the full story of gene silencing would be a mammoth enterprise that would take me many years to write and would take you well into the night to read. So we’ll need to abbreviate the story more than a little. Second (and as you will see) we are only in the dawn of our knowledge; so consider the following to be primer... the best we could do as of December 8th, 2006. And third, please understand that the story that I am telling represents the work of several generations of biologists, chemists, and many shades in between. I’m pleased and proud that work from my labo- ratory has contributed to the field, and that this has led to my being chosen as one of the messengers to relay the story in this forum. At the same time, I hope that there will be no confusion of equating our modest contributions with those of the much grander RNAi enterprise. DOUBLE STRANDED RNA AS A BIOLOGICAL ALARM SIGNAL These disclaimers in hand, the story can now start with a biography of the first main character. Double stranded RNA is probably as old (or almost as old) as life on earth.
    [Show full text]
  • Clinical Molecular Genetics in the Uk C.1975–C.2000
    CLINICAL MOLECULAR GENETICS IN THE UK c.1975–c.2000 The transcript of a Witness Seminar held by the History of Modern Biomedicine Research Group, Queen Mary, University of London, on 5 February 2013 Edited by E M Jones and E M Tansey Volume 48 2014 ©The Trustee of the Wellcome Trust, London, 2014 First published by Queen Mary, University of London, 2014 The History of Modern Biomedicine Research Group is funded by the Wellcome Trust, which is a registered charity, no. 210183. ISBN 978 0 90223 888 6 All volumes are freely available online at www.history.qmul.ac.uk/research/modbiomed/ wellcome_witnesses/ Please cite as: Jones E M, Tansey E M. (eds) (2014) Clinical Molecular Genetics in the UK c.1975–c.2000. Wellcome Witnesses to Contemporary Medicine, vol. 48. London: Queen Mary, University of London. CONTENTS What is a Witness Seminar? v Acknowledgements E M Tansey and E M Jones vii Illustrations and credits ix Abbreviations xi Ancillary guides xiii Introduction Professor Bob Williamson xv Transcript Edited by E M Jones and E M Tansey 1 Appendix 1 Photograph, with key, of delegates attending The Molecular Biology of Thalassaemia conference in Kolimbari, Crete, 1978 88 Appendix 2 Extracts from the University of Leiden postgraduate course Restriction Fragment Length Polymorphisms and Human Genetics, 1982 91 Appendix 3 Archival material of the Clinical Molecular Genetics Society 95 Biographical notes 101 References 113 Index 131 Witness Seminars: Meetings and Publications 143 WHAT IS A WITNESS SEMINAR? The Witness Seminar is a specialized form of oral history, where several individuals associated with a particular set of circumstances or events are invited to meet together to discuss, debate, and agree or disagree about their memories.
    [Show full text]
  • Journal of Biomedical Discovery and Collaboration
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE Journal of Biomedical Discovery and provided by PubMed Central Collaboration BioMed Central Case Study Open Access The emergence and diffusion of DNA microarray technology Tim Lenoir*† and Eric Giannella† Address: Jenkins Collaboratory for New Technologies in Society, Duke University, John Hope Franklin Center, 2204 Erwin Road, Durham, North Carolina 27708-0402, USA Email: Tim Lenoir* - [email protected]; Eric Giannella - [email protected] * Corresponding author †Equal contributors Published: 22 August 2006 Received: 09 August 2006 Accepted: 22 August 2006 Journal of Biomedical Discovery and Collaboration 2006, 1:11 doi:10.1186/1747-5333-1-11 This article is available from: http://www.j-biomed-discovery.com/content/1/1/11 © 2006 Lenoir and Giannella; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract The network model of innovation widely adopted among researchers in the economics of science and technology posits relatively porous boundaries between firms and academic research programs and a bi-directional flow of inventions, personnel, and tacit knowledge between sites of university and industry innovation. Moreover, the model suggests that these bi-directional flows should be considered as mutual stimulation of research and invention in both industry and academe, operating as a positive feedback loop. One side of this bi-directional flow – namely; the flow of inventions into industry through the licensing of university-based technologies – has been well studied; but the reverse phenomenon of the stimulation of university research through the absorption of new directions emanating from industry has yet to be investigated in much detail.
    [Show full text]
  • Blotting Techniques
    BLOTTING TECHNIQUES Dr.Ramesh C.K. Associate Professor and Chairman Department of Biotechnology Sahyadri Science College Shimoga - 577 203 Karnataka, India BLOTTING TECHNIQUES Southern Blot Northern blot Western blot It is used to detect It is used to detect It is used to detect the DNA. the RNA. proteins. SOUTHERN BLOTTING A technique for identifying specific sequences of DNA in which DNA fragments are separated by electrophoresis, transferred to a membrane, and identified with a suitable probe. Detects restriction fragments following a restriction enzyme digest of genomic DNA (RFLPs) Named after British biochemist Edwin Southern (alive and publishing!) Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol Biol. 1975 Nov 5;98(3):503-17. History/Background • ‘Southern’ hybridization named after Sir Edwin Southern • Developed in 1975 • One of the most highly cited scientific publications “Used to detect the DNA” • This method Involves: Separation Transfer Hybridization. • This DNA can be: • Single gene • Part of a larger piece of DNA……..viral genome “The key to this method is Hybridization” Hybridization “Process of forming a dsDNA molecule between a ssDNA probe and a ss-target DNA” PRINCIPLE The mixture of molecules is separated. Immobilized on a matrix. Probe addition to the matrix to bind to the molecules. Unbound probes are removed. “The place where the probe is connected corresponds to the location of the immobilized target molecule.” Steps The DNA is digested Fragments Gel electrophoresis Transfer to membrane Probing Autoradiogram I. DNA Purification – Isolate the DNA in question from the rest of the cellular material in the nucleus.
    [Show full text]