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Recombinant_DNA_Technology_Class12.pptxok
- 1. Recombinant DNA Technology •Class 12 Biology / Biotechnology • Detailed PPT Presentation
- 2. Introduction to rDNA •Recombinant DNA (rDNA) technology is a technique to join DNA from two different sources. • The resulting DNA is inserted into a host organism to express the desired gene. • Used widely in medicine, agriculture, research, and industry.
- 3. Tools of rDNATechnology • - Restriction Enzymes: Cut DNA at specific sites. • - DNA Ligase: Joins DNA fragments. • - Vectors: Carry foreign DNA (e.g., plasmids, viruses). • - Host Cells: Organisms into which recombinant DNA is inserted. • - Selectable Markers: Help identify transformed cells.
- 4. Steps in rDNATechnology – Overview • 1. Isolation of genetic material (DNA) • 2. Cutting DNA at specific sites using restriction enzymes • 3. Amplification of gene of interest using PCR • 4. Insertion of DNA into vector • 5. Introduction into host • 6. Selection of recombinants • 7. Expression and purification of product
- 5. Isolation of DNA& Restriction Digestion • DNA is extracted from donor and vector cells. • Restriction enzymes cut DNA at specific sequences to obtain target gene and prepare vector.
- 6. PCR – Amplificationof Gene of Interest • Polymerase Chain Reaction (PCR) makes millions of copies of the gene. • Steps: • 1. Denaturation • 2. Annealing • 3. Extension • Components: DNA template, primers, DNA polymerase, dNTPs.
- 7. Ligation – JoiningDNA with Vector • Cut vector and gene are joined using DNA ligase. • Forms recombinant DNA molecule.
- 8. Introduction into HostCell • The recombinant plasmid is introduced into a host cell (e.g., E. coli). • Techniques: Heat Shock, Electroporation, Microinjection, Gene Gun.
- 9. Selection and Screeningof Recombinants • Not all host cells take up recombinant DNA. • Selectable markers like antibiotic resistance are used. • Only transformed cells grow on selective media.
- 10. Expression and Purificationof Gene Product • Host cell expresses the inserted gene to produce protein. • The protein is then extracted and purified. • Used for medicine, research, etc.
- 11. Applications of rDNATechnology • - Medicine: Insulin, vaccines, gene therapy • - Agriculture: GM crops (Bt cotton, Golden Rice) • - Industry: Enzymes (amylase, cellulase) • - Environment: Bioremediation, oil spill treatment
- 12. Conclusion • Recombinant DNAtechnology has revolutionized biotechnology. • It enables targeted genetic manipulation for beneficial outcomes. • Its applications continue to grow in every field of life sciences.