lecture-recombinant-DNA-technology....ppt
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The document discusses recombinant DNA technology, outlining its vocabulary, procedures, and applications. It details the manipulation of DNA from different species, creation of recombinant molecules, and the use of vectors for gene transfer. Applications include medical advancements, genetic testing, and forensic use, highlighting its significance in various fields such as medicine, agriculture, and research.
lecture-recombinant-DNA-technology....ppt
- 1. Dr SARITA SHARMA ASSOCIATE PROFESSOR MMCP, MMDU Recombinant DNA technology
- 2. Essential vocabulary Biotechnology/ GE RDNA/ RDNA tech Restriction enzymes/endonucleases Cloning DNA ligase Vectors Plasmid Bacteriophages DNA library
- 3. Biotechnology The manipulation of biological processes and/or organisms for the benefit of mankind .
- 4. RDNA DNA that has been created artificially (not natural). DNA from two or more sources is incorporated into a single recombinant molecule
- 5. RDNA tech Procedures by which DNA from different species can be isolated, cut and spliced together New "recombinant " molecules are then multiplied in quantity in populations of rapidly dividing cells (e.g. bacteria, yeast). Uses methods derived from biochemistry of nucleic acids coupled with genetic techniques originally used for the study of bacteria and viruses
- 7. RDNA tech process 1. A gene is located on a chromosome map (RFLP) 2. A DNA library is constructed – plasmid are obtained, cleavage occurs and RDNA prod. 3. The gene of interest is isolated (cloned) from the library via plasmid DNA isolation, restriction digestion and electrophoresis. 4. Multiple copies of gene interest are produced for study.
- 8. steps Restriction enzymes “cuts” DNA at specific sites (restriction fragments) DNA ligase “pastes” the DNA fragments together The cut segments are inserted into other DNA The cut segments are inserted into other DNA molecules that serves as molecules that serves as vectors
- 9. Vectors Carrier DNA mol = transfers the RDNA into the host cell. Within host cells, vectors can replicate producing many DNA segments = identical copies (CLONES) Host cells pass the recombinant DNA mol on their progeny = population of cells. Cloned DNA segments recovered from host cells for purification and analysis
- 10. Plasmids Molecules of DNA that are found in bacteria Act as a system to transfer genetic material to other bacteria, allowing those to express the transmitted genes. small (a few thousand base pairs) usually carry only one or a few genes circular have a single origin of replication
- 11. Resctriction enzymes DNA cutting enzymes. Restriction endonucleases cuts DNA at a specific site defined by a sequence of bases in the DNA (recog.site) forming “sticky ends” ( Palindromic sites several hundred endonucleases have been extracted from bacteria and many are used in recombinant DNA research. eg EcoR1,Hind III, HaeIII, TaqA1, Sau3A
- 13. Library construction Because each cloned DNA segemnt is relatively small, many separate clones must be constructed . A set of cloned DNA segments derived from a single individual represents a library Cloned libraries could be an entire genome, a singe chromosome, or a set og genes compiled together in a single cell type.
- 15. summary
- 16. Applications of recombinant DNA Applications of recombinant DNA technology technology Used widely in research and hospital laboratories. Used widely in research and hospital laboratories. Broad applications - medicine, agriculture, Broad applications - medicine, agriculture,
- 17. ??.. 1. The processes of inheritance and gene expression. 2. Process of various diseases 3. Generation of economic benefits eg. improved agricultural products.
- 18. Applications Molecular Biology/Research Diagnostics Genetic Counseling Criminology/Forensics Paternity testing Archeology Food testing Evolutionary studies
- 20. Medicine Medicine –production of industrial and commercial compounds Insulin – Diabetes drugs (angiostation and endostatin) , Factor VIII – Haemophilia A Factor IX – Haemophilia B EPO – Anaemia Interleukins and interferons Tissue plasminogen activator – dissolve blood clots Hormones = GH, parathyroid Oxytocin Adenosine deaminase –sev. Com. Imm (SCID) ,
- 21. Diagnostic kits Diagnostic kits – Hep, AIDS Antibiotics Antibiotics genetic testing,–Mapping the chromosomal location of genetic disorders. RFLP, DNA fingerprinting – the HG project Gene therapy – manipulation of DNA to tx diseases by altering individuals genes Forensic applications - All individuals are genetically unique = a distinct "genetic fingerprint“, all types of specs, old and new Animals = models of genetic diseases Eg mice