Gene Cloning
CBSE · Class 12 · Biotechnology
Complete topic list for Gene Cloning in CBSE Class 12 Biotechnology. Key concepts, sub-topics, and what to focus on for board exams.
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3.1 Identification of Candidate Gene
- The first and most formidable step in gene cloning is identifying the candidate gene of interest.
- Candidate genes are identified based on their biomedical, economic, or evolutionary significance.
- Examples: Insulin gene (diabetes), iron-uptake gene (chlorosis), salinity-tolerance gene, milk protein genes, blood-clotting factor genes.
3.2 Isolation of Nucleic Acids
- Two major challenges: (i) Nucleic acids are present in very small amounts compared to proteins, carbohydrates and lipids; (ii) Large size of DNA makes it susceptible to cleavage under physical stress.
- Four steps in nucleic acid extraction: (1) Cell disruption/lysis, (2) Protection from degrading nucleases, (3) Separation from other molecules, (4) Precipitation and concentration using ethanol or iso
- Bacterial cells: Lysozyme digests peptidoglycan cell wall; SDS (anionic detergent) lyses cell membrane.
3.3 Enzymes Used for Recombinant DNA Technology
- Enzymes are the 'molecular scissors and glue' of rDNA technology.
- NUCLEASES: Cleave nucleic acids by hydrolysing phosphodiester bonds. Types: (i) Exonucleases — remove mononucleotides from 3′ or 5′ ends; (ii) Endonucleases — cleave internal phosphodiester bonds with
- RESTRICTION ENDONUCLEASES (REs): Endonucleases that recognise and cleave at specific palindromic sequences (4–8 bp). Mostly found in bacteria and archaea as defence against bacteriophages.
3.4 Modes of DNA Transfer
- Transfer of foreign DNA into a host cell is a key step in rDNA technology.
- NATURAL METHODS in bacteria: (i) Transformation, (ii) Transduction, (iii) Conjugation.
- TRANSFORMATION: Direct uptake of exogenous DNA from surroundings through the cell membrane; recipient cells are called transformants. Occurs naturally in some bacteria.
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Central concept: Gene Cloning
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Sources & Official References
- NCERT Official — ncert.nic.in
- CBSE Academic — cbseacademic.nic.in
- CBSE Official — cbse.gov.in
- National Education Policy 2020 — education.gov.in
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