BIOTECHNOLOGY (878)

CLASS XII There will be two papers in the subject: (ii) Protein Synthesis: synthesis of different RNAs, and the complete mechanism of Paper I: Theory……………. 3 hours ... 70 marks polypeptide chain formation. Paper II: Practical…………. 3 hours ... 15 marks Concept of central dogma. Project Work………. … 10 marks From genes to proteins: Practical File………… … 5 marks (a) Concept of transcriptional unit, promoter, structural and terminator region; concept PAPER I: THEORY- 70 Marks of split gene - intron and exon; hnRNA; There will be one paper of three hours duration (b) Transcription – explanation of the divided into two parts. complete process including enzymes Part 1 (20 marks) will consist of compulsory short involved in the process; Post- answer questions, testing knowledge, application and transcriptional changes and their skills relating to elementary/fundamental aspects of significance in eukaryotes – the entire syllabus. polyadenylation, capping and RNA Part 2 (50 marks) will consist of eight questions out splicing; of which the candidates will be required to answer (c) Concept of reverse transcription; five questions. Each question in this part shall carry (d) Genetic code – properties of genetic code, 10 marks. start and stop codons, anticodons. 1. Molecular Biology (e) The translation of RNA to protein – (i) Nucleic acids and their estimation: an complete mechanism of chain initiation, understanding of nucleic acids, their elongation and termination, the role of biochemical structure. tRNA, mRNA and rRNA in protein DNA as the genetic material (Hershey and synthesis. (Post translational changes not Chase experiment). included). DNA (B-DNA)– physical and chemical (iii) Gene regulation in prokaryotes structure; definition, double helical model of Operon concept – lac operon. DNA, (Watson and Crick’s); Nucleotide and nucleoside; Chargaff’s Law, method of 2. Genetic Engineering replication of DNA, various replicative (i) Introduction to gene cloning and genetic enzymes in both prokaryotic and eukaryotic engineering: concept of cloning and vectors. organisms, example topoisomerases, helicase, Tools of recombinant DNA technology, types of SSBs polymerases, primases, ligases. Concept restriction endonucleases and other enzymes of semi conservative (with respect to used in gene cloning; techniques involved in Messelson and Stahl experiment and semi- extraction and purification of DNA from discontinuous replication, (leading and bacterial and plant cells. lagging strands), okazaki fragments. Selection of host cells: eukaryotic and RNA – definition, various types of RNAs such prokaryotic. as mRNA, tRNA (Clover leaf model with diagram; rRNA their structure and functions. Vectors: Characteristics and types such as plasmids -pBR322, pUC (in pBR322- presence Technique of nucleic acid estimation – of two antibiotic resistant genes and in colorimetry. pUCpresence of lac Z gene to be taught), 1

cosmids, phages (M13 and λ), YACs, BACs (to the technique to be taught), western and be taught with reference to stability and their northern blotting (a brief idea and their uses). carrying capacity), animal and plant viruses High resolution techniques: DNA sequencing- (CaMV, retrovirus, SV40 – only names of sequencing by chain termination. Site directed viruses, no details). mutagenesis. Transfer of recombinants into host cells – DNA amplification by Polymerase chain (a) Vectorless methods - basic concept of reaction (PCR)– applications of PCR, steps transformation, transfection, and application of DNA profiling or DNA electroporation, liposome mediated gene finger printing. transfer, microinjection, biolistic 3. technology (b) Vector-mediated method - Agrobacterium tumefaciens induced gene transfer. A brief idea of tools and techniques involved in cell culture technology and their applications in Methods of identification of recombinants- microbial, plant tissue and animal cell cultures Direct selection (green fluorescent respectively. selection) and Insertional inactivation (Blue-white selection). (i) General tools and techniques used in cell culture technology A basic understanding of DNA libraries – construction of genomic and cDNA (a) Instruments - CO2 , bioreactor libraries. (diagram, its components and their function)-stirred tank and sparged type Construction of a recombinant DNA (brief idea only), use of T flasks to molecule. propagate animal cells. (ii) Innovations in Biotechnology: produced by Only uses of the above instruments to using modern biotechnological tools, (select be studied. examples of products already available) (b) Sterilization techniques for culture room, (a) Plants: Production of Flavr Savr apparatus, transfer area, media, tomatoes and Golden rice. vitamins, and living material; (b) Healthcare: Production of recombinant (c) Cryopreservation (need and steps). hepatitis-B vaccine, Humulin and (d) Cell counting (direct counting by interferon. haemocytometer), cell viability by (c) Animal: Dolly the cloned sheep, Sources Evan’s blue stain and cell sorting (FACS and characteristics of stem cells and only) their applications. (e) Types of media (synthetic /defined, semi- synthetic/differential, complex/natural) (d) Environmental biotechnology: bioremediation using oil-eating bacteria Preparation of media: microbial media- as an example. LB agar; Plant media-MS and White’s (e) Industrial biotechnology: applications of media; Animal media-RPMI - brief idea industrial enzymes – rennet, subtilisin, only. (includes inorganic and organic amylase, papain. macronutrients and micronutrients, (iii) Gene analysis techniques: various techniques antibiotics, growth regulators for plants: involved in recombinant DNA technology. auxins and cytokinins). DNA probes – definition and use. Importance of pH and solidifying agents. Low resolution mapping techniques: gel electrophoresis, southern blotting (details of 2

(ii) Microbial culture and its application. Data retrieval tools- ENTREZ, Taxonomy Browser. Fermentation process and growth kinetics- batch culture, fed batch culture, continuous (ii) Genomics: Definition, introduction, tools used culture (with the help of graphs only): in Genomics and its applications. Definition of and : Definition of genomics. Types of genomics- Products and application-SCP (definition and structural and functional. use). Types of sequence analysis by using BLAST and FASTA. (iii) Plant tissue culture and its application. Human Genome Project - its objectives, the Isolation of single cell by mechanical and countries involved, its achievements and enzymatic methods, synchronisation of cell significance. culture by chemical methods like starvation, (iii) Proteomics: definition, introduction and inhibition and mitotic arrest. databases. Cellular totipotency-definition of cellular Types of Proteomics – structural and differentiation, de-differentiation, re- functional. differentiation. Application of plant cell

culture technology (methodology not required, only brief idea needed): PAPER II (a) Haploid production-androgenesis and PRACTICAL WORK – 15 marks gynogenesis and their significance. Candidates are required to complete the following (b) Triploid production-understanding and experiments. need for triploid production and its 1. Paper Chromatography – separation of application (seedless crops). photosynthetic pigments (c) In-vitro pollination- concept and its Take any leaf. Extract chlorophyll in 80% application. acetone. Take a strip of paper or prepare a thin (d) Somatic hybridisation-protoplast fusion layer of silica gel on a slide. Load chlorophyll (Pomato). extract at one end of the paper/gel. Keep paper (e) Micropropagation and its significance. or gel in the rising medium in or jar for (f) Biodegradable plastics (concept of PHB). about 30 minutes. The rising medium should have methanol/ acetic acid, n-butanol or benzene. The (iv) Animal cell culture and its application. rising fluid should always be at the bottom below Primary cell culture with enzymatic the point of loading of chlorophylls. After 30 disaggregation and its drawbacks; Types of minutes, three spots: yellow, bluish green and cell-lines: finite, continuous, adherent and light green will be observed corresponding to suspension; scale up-mono layer by Roller carotenes, chlorophyll A & chlorophyll B. , application of animal cell culture- tissue, hybridoma technology, tissue 2. Preparation of buffers – acetate buffers engineering (definition only). This experiment should be done to make the basics clear to the students. Basic calculation for 4. Bioinformatics buffer preparation should be known. The (i) Introduction to bioinformatics; global approach should be to utilize easily available bioinformatics databases and data retrieval chemicals at reasonable costs. Acetate buffers can tools; genomics, different types of sequences, give the range of pH 4 - pH 9.2 types of sequence analysis. Introduction to bioinformatics: definition and 3. Sterilization of culture medium and other need. materials. An introduction to global bioinformatics (i) Dry Physical method – heat or radiation. databases (nucleotide and protein databases). Information sources such as EMBL, NCBI, (ii) Wet Physical methods – steam sterilization. DDBJ, SWISSPROT, GenBank, GENSCAN. 3

(iii) Chemical Sterilization/ Surface sterilization PROJECT WORK AND PRACTICAL FILE Disinfection with 70% alcohol and Sodium hypochlorite solution carbolic acid – 15 Marks 4. Preparation of various forms of culture media – Project Work – 10 Marks Petri plate, slant and suspension. Luria Bertani (L.B) media to be prepared, The Project Work is to be assessed by a Visiting Examiner appointed locally and approved by the autoclaved and cooled to 60 degrees C. To Council. prepare nutrient plates the media is poured into presterilized petri-dishes under a LAF. To Candidates are to creatively execute one prepare slants the media is poured into several project / assignment on an aspect of Biotechnology. test tubes, plugged and kept in a tilted position (at an angle of 45o) until it sets. Teachers may assign or students may choose any one project of their choice. The report should be kept 5. Identification of bacteria by Gram +ve and Gram simple, but neat and elegant. No extra credit shall be –ve (from curd /saliva and/or soil solution) given for type-written material/decorative cover, etc. (i) Prepare a bacterial smear on a slide (ii) Stain with crystal violet stain. (iii) Rinse with water. A list of suggested projects is as follows: (iv) Add a few drops of iodine solution. (v) Add 1. Effluent analysis. few drops of 90 % ethanol (vi) Counterstain with 2. A study of the technological details of malt safranin solution (vii) Observe the red and blue preparation. colonies under the 3. A study of the technological details of the 6. Action of enzymes on starch under: (a) variable brewing industry. temperature (b) variable substrate concentration – 4. A study of the organisation of a fermenter. plotting of Km value by graph 5. Technological analysis of the process of drug Soluble starch solution (0.5% - 1%) to be development, drug designing and drug targeting. prepared. Test with iodine. Collect saliva, dilute 6. A study of the technological details of vaccine 1: 5, add 1 ml of saliva to 10 ml of starch solution. development. Incubate for 15 minutes. Again test for presence 7. Diagnosis of diseases by modern techniques like of starch with iodine. Also test for the presence of ELISA, RIA and Antibody targeting. reducing sugars in solution. Repeat the same 8. DNA finger-printing. process at the variable volumes of starch. 9. DNA foot-printing. 7. Isolation of DNA from plants 10. Microbiological contaminants in food and food Take half a ripe and peeled banana into a beaker products. and add 50 ml of extraction fluid (1.5gm table salt 11. Isolation of microbes from air, water and soil. +10 ml liquid detergent +90 ml distilled water). Place the beaker in a water bath set at 60 degrees 12. Methods of identifying microbes (various C for 15 minutes. Stir gently with a . staining techniques and biochemical reactions). Filter 5ml of cooled content into a clean test tube 13. Tissue Culture and its applications. and add 5ml of cold 90% ethanol. DNA molecules 14. Stem Cell Technology separate out and appear as white fibres. [DNA 15. Nanotechnology can also be extracted from pea seeds and soaked 16. Bioinformatics wheat grains] 17. Genetic Engineering 8. Isolation of milk protein – wet weight and dry 18. Cloning weight. 19. Instrumentation in biotechnology Milk proteins are isolated by adding 0.4 N HCl 20. Forensic Biotechnology into the milk sample. Casein start coagulating at 21. Ethical, Legal and Social Issues (ELSI) related to its isoelectric point (i.e. at pH 4.6). The Biotechnology/ GMOs precipitate is filtered and weighed to quantify the 22. Biopiracy- Case Studies protein present.

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Practical File – 5 Marks – Presentation (graphs, tables, charts, newspaper The Visiting Examiner is required to assess students cuttings, handmade diagrams, photographs, on the basis of the practical file maintained by them statistical analysis if relevant) during the academic year. – Conclusion/ Summary – Bibliography Suggested Evaluation Criteria for Project Work: Projects should be handwritten by the candidate. Format of the Project: Written pages should not exceed 15-20 pages. – Content – Introduction

LIST OF EQUIPMENT FOR BIOTECHNOLOGY PRACTICALS FOR CLASSES XI & XII 1. Table-top Centrifuge 12. Incubator 2. Vortex - Mixer 13. with 3. Thermostatic water-bath 14. Laminar flow cabinet 4. Spectrophotometer (UV visible range)/ 15. Weighing Balance (Electrical) 16. Hot plate 5. Refrigerator 17. Binocular Microscope 6. Lactometer 18. Haemocytometer 7. pH meter 19. Colony counter 8. 20. Antiserum 9. 21. Electrophoresis chamber 10. Desiccators 22. Micropipettes 11. Micro-filtration unit

LIST OF ABBREVIATIONS TO BE STUDIED 1. BAC: Bacterial Artificial Chromosomes 15. ICGEB: International Centre for Genetic 2. BLAST: Basic Local Alignment Search Tool Engineering and Biotechnology 3. CTAB: Cetyl Trimethyl Ammonium Bromide 16. IFN: Interferon 4. DBM: Diazo–benzyl oxy–methyl paper 17. LB medium: Luria and Bertani Medium 5. DDBJ: DNA Database/ Data Bank of Japan 18. MS medium: Murashige and Skoog medium 6. ddNTP: Dideoxy Nucleoside triphosphate 19. NCBI: National Centre for Biotechnology 7. EBI: European Bioinformatics Institute Information 8. EMBL: European Molecular Biology 20. NHGRI: National Human Genome Research 9. EST: Expressed Sequence Tag Institute 10. FACS: Fluorescence Activated Cell Sorting 21. PAGE: Polyacrylamide Gel Electrophoresis 11. FASTA: Fast All 22. PCR: Polymerization Chain Reaction 12. HEPA: High Energy Particulate Air 23. PDB: Protein Database/ Data Bank 13. HGP: Human Genome Project 24. PHB: Poly 3–Hydroxyl Butyrate 14. IBPGR: International Board of Plant Genetic 25. PIR: Protein Information Resource Resources 26. RFLP: Restriction Fragment Length Polymorphism

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27. RNA: Ribonucleic acid 28. RPMI medium: Roswell Park Memorial Institute medium 29. SCP: Single Cell Protein 30. SDS – PAGE: Sodium Dodecyl Sulphate– Polyacrylamide Gel Electrophoresis 31. SNP: Single Nucleotide Polymorphism 32. SSBs: Single Stranded Binding Proteins 33. STS: Sequence Tagged Site 34. VNTR: Variable Number of Tandem Repeats 35. YAC: Yeast Artificial Chromosome

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