Recombinant Dna Technology
BIOTECHNOLOGY
RECOMBINANT DNA technology 1. This is a modern biotechnological advance , in which a desired gene fragment can be inserted in to a cloning vector and the resulting DNA (Recombinant DNA) can be amplified in suitable host. 2. A vector can be a plasmid, cosmid,bacterophage,retroviruses, animal and plant viruses or artificial chromosomes like YAC, BAC,or HAC.(Yeast artificial chromosome, bacterial........) 3. The rec. DNA produced can be amplified or cloned in a suitable vector like bacteria for plamids, cosmids or bacterophages, plant and animal cells for viruses .
Involves five steps:
1. Enzyme restriction digest of DNA sample. 2. Enzyme restriction digest of DNA plasmid vector (same Res.Enzyme).
3. Ligation of DNA sample products and plasmid vector.
4. Transformation with the ligation products.
5. Growth on agar plates with selection for antibiotic resistance.
[pic]
← APPLICATION OF RECOMBINANT DNA technology ← Gene therapy ← Recombinant Vaccines ← Genetically modified crops ← Biosensors ← Monoclonal antibodies ← Cell/tissue culture ← Xenotransplantation ← Bioremediation ← Production of next generation antibiotics ← Forensics ← Bioterrorism detection
Cloning vectors DNA molecules that are used to "transport" cloned sequences between biological hosts and the test tube. Cloning vectors share four common properties: 1. Ability to promote autonomous replication. 2. Contain a genetic marker (usually dominant) for selection. 3. Unique restriction sites to facilitate cloning of insert DNA. 4. Minimum amount of nonessential DNA to optimize cloning.
Main types of cloning vectors
Plasmid, bacteriophage, cosmid, bacterial artificial chromosome (BAC), yeast artificial chromosome (YAC), HAC, retrovirus, baculovirus vector etc.
1. PLASMIDS ← Bacterial cells may contain extra-chromosomal DNA called plasmids. ← Plasmids are usually
RECOMBINANT DNA technology 1. This is a modern biotechnological advance , in which a desired gene fragment can be inserted in to a cloning vector and the resulting DNA (Recombinant DNA) can be amplified in suitable host. 2. A vector can be a plasmid, cosmid,bacterophage,retroviruses, animal and plant viruses or artificial chromosomes like YAC, BAC,or HAC.(Yeast artificial chromosome, bacterial........) 3. The rec. DNA produced can be amplified or cloned in a suitable vector like bacteria for plamids, cosmids or bacterophages, plant and animal cells for viruses .
Involves five steps:
1. Enzyme restriction digest of DNA sample. 2. Enzyme restriction digest of DNA plasmid vector (same Res.Enzyme).
3. Ligation of DNA sample products and plasmid vector.
4. Transformation with the ligation products.
5. Growth on agar plates with selection for antibiotic resistance.
[pic]
← APPLICATION OF RECOMBINANT DNA technology ← Gene therapy ← Recombinant Vaccines ← Genetically modified crops ← Biosensors ← Monoclonal antibodies ← Cell/tissue culture ← Xenotransplantation ← Bioremediation ← Production of next generation antibiotics ← Forensics ← Bioterrorism detection
Cloning vectors DNA molecules that are used to "transport" cloned sequences between biological hosts and the test tube. Cloning vectors share four common properties: 1. Ability to promote autonomous replication. 2. Contain a genetic marker (usually dominant) for selection. 3. Unique restriction sites to facilitate cloning of insert DNA. 4. Minimum amount of nonessential DNA to optimize cloning.
Main types of cloning vectors
Plasmid, bacteriophage, cosmid, bacterial artificial chromosome (BAC), yeast artificial chromosome (YAC), HAC, retrovirus, baculovirus vector etc.
1. PLASMIDS ← Bacterial cells may contain extra-chromosomal DNA called plasmids. ← Plasmids are usually