Recombinant DNA Technology
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Recombinant DNA Technology Method
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Recombinant DNA Technology
- 1. Recombinant DNA Technology Presented By Muhammad Aqib Shabbir Bachelor of Science in Biotechnology Semester 5th Course Instructor Dr. Sajid Mansoor
- 2. Contents • Introduction • Discovery • Goals & Objectives • rDNA Technology Procedure • Enzymes • Vectors • Techniques • Applications
- 3. Introduction • DNA Molecules that are extracted from different sources and chemically joined together • For Example DNA comprising an animal gene may be recombined with a DNA from bacterium
- 4. Discovery Discovery of DNA Structure Watson & Crick in 1953 Isolation of DNA ligase in 1967 Isolation of REase in 1970 Paul Berg generated rDNA technology in 1972
- 5. Goals of rDNA Technology • To isolate and characterize a gene • To make desired alterations in one or more isolated genes • To return altered genes to living cells • Artificially synthesize new gene • Alternating the genome of an organism • Understanding the hereditary diseases and their cure • Improving human genome
- 6. Procedure Isolating of DNA • Extraction of DNA Cutting of DNA • DNA can be cut into large fragments by mechanical shearing • Restriction enzymes are the scissors of molecular genetics
- 7. Joining of DNA • DNA Ligase Amplifying of DNA • DNA Clone
- 10. Joining of DNA
- 12. Enzymes Enzyme Role DNA Ligase Bind to DNA Molecule Type II Restriction Endonucleases Cleaves DNA at specific sites Reverse Transcriptase Make a DNA copy of RNA Molecule DNA Polymerase I Fill single stranded gaps of DNA duplex Polynucleotide Kinase Adds a Phosphate to 5’-OH end of Polynucleotide Terminal Transferase Adds Homopolymer tails to 3’-OH ends Exonuclease III Removes nucleotide residues from the 3’ ends Bacteriophage Exonuclease Removes nucleotides from 5’ ends Alkaline Phosphatase Removes terminal phosphates
- 14. Techniques • Gel Electrophoresis • Cloning Libraries • Restriction Enzyme Mapping • PCR • Nucleic Acid Hybridization • DNA Microarrays
- 15. Application • Agriculture: Growing crops of your choices (GM Food) • Pharmacology: Artificial Insulin, Drug delivery to target sites • Medicine: Gene Therapy, Antiviral Therapy • Other Uses: Fluorescent Fishes, Glowing Plants etc.
- 16. ?