PPT On Recombinant DNA
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2. What is recombinant DNA?
Recombinant DNA is an artificial strand of DNA created by combining DNA segments from two or more sources. It is the basis for all sorts of genetic engineering.
3. Uses of recombinant DNA
Genetically modified crops (Roundup Ready) Genetically modified animals for research (Oncomouse) Genetically modified micro-organisms to produce pharmaceuticals (insulin) Gene therapy (Cystic Fibrosis) To isolate and study a particular gene (subcloning) Creating transgenic organisms or chimeras.
4. In vitro recombination A restriction enzyme “cuts” both DNA strands. The cut creates single-stranded sections known as the “sticky ends.” If cut with the same enzyme, they are complimentary. Complimentary sticky ends pair with each other. DNA ligase “seals” the two strands together, forming one strand of DNA. Once created, the recombinant DNA must be replicated either in vitro through polymerase chain reaction (PCR) or inside various cells.
5. Marker genes
A marker gene, which produces a distinctive phenotype is added to the genome. It is necessary to identify recombinant organisms from unaffected organisms. For example, the gene for ampicillin resistance (an antibiotic) is often added to the bacterial genome. When the culture is treated with ampicillin, only the recombinant bacteria remain. Other marker genes, usually for eukaryotes, include fluorescence (a gene from jellyfish).
6. Transformation methods
Microinjection: a needle directly injects the DNA into nuclei of embryonic cells. Particle bombardment: tiny gold or tungesten particles are coated w/ DNA and shot into cells. Administered with a gene gun. Electroporation: cells are shocked, creating holes in membrane for DNA to enter. Transformation is the process of a cell altering its genome by uptake of foreign DNA. Recombinant DNA must be inserted into host organism to be of use.
7. Cloning Vectors
The recombinant DNA sequence can also be introduced into the host by a vector. For bacteria: Plasmids—round extra-chromosomal self-replicating DNA Bacteriophages—viruses that infect bacteria The most efficient vectors for uptake of DNA into eukaryotes are viruses: Retroviruses—single RNA strand infects by reverse transcription, where RNA is transcribed into DNA. Adenoviruses—able to infect most cells with no toxic effects. Parvoviruses—suppresses cancer genes; used for gene therapy against tumors. Herpesviruses—able to infect many types of cells, including neurons. Poxviruses—triggers strong immune response. Used against tumors for cancer gene therapy.
8. For More Please Refer Our PPT. Thank You.
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Presentation Transcript:
1. Recombinant DNA2. What is recombinant DNA?
Recombinant DNA is an artificial strand of DNA created by combining DNA segments from two or more sources. It is the basis for all sorts of genetic engineering.
3. Uses of recombinant DNA
Genetically modified crops (Roundup Ready) Genetically modified animals for research (Oncomouse) Genetically modified micro-organisms to produce pharmaceuticals (insulin) Gene therapy (Cystic Fibrosis) To isolate and study a particular gene (subcloning) Creating transgenic organisms or chimeras.
4. In vitro recombination A restriction enzyme “cuts” both DNA strands. The cut creates single-stranded sections known as the “sticky ends.” If cut with the same enzyme, they are complimentary. Complimentary sticky ends pair with each other. DNA ligase “seals” the two strands together, forming one strand of DNA. Once created, the recombinant DNA must be replicated either in vitro through polymerase chain reaction (PCR) or inside various cells.
5. Marker genes
A marker gene, which produces a distinctive phenotype is added to the genome. It is necessary to identify recombinant organisms from unaffected organisms. For example, the gene for ampicillin resistance (an antibiotic) is often added to the bacterial genome. When the culture is treated with ampicillin, only the recombinant bacteria remain. Other marker genes, usually for eukaryotes, include fluorescence (a gene from jellyfish).
6. Transformation methods
Microinjection: a needle directly injects the DNA into nuclei of embryonic cells. Particle bombardment: tiny gold or tungesten particles are coated w/ DNA and shot into cells. Administered with a gene gun. Electroporation: cells are shocked, creating holes in membrane for DNA to enter. Transformation is the process of a cell altering its genome by uptake of foreign DNA. Recombinant DNA must be inserted into host organism to be of use.
7. Cloning Vectors
The recombinant DNA sequence can also be introduced into the host by a vector. For bacteria: Plasmids—round extra-chromosomal self-replicating DNA Bacteriophages—viruses that infect bacteria The most efficient vectors for uptake of DNA into eukaryotes are viruses: Retroviruses—single RNA strand infects by reverse transcription, where RNA is transcribed into DNA. Adenoviruses—able to infect most cells with no toxic effects. Parvoviruses—suppresses cancer genes; used for gene therapy against tumors. Herpesviruses—able to infect many types of cells, including neurons. Poxviruses—triggers strong immune response. Used against tumors for cancer gene therapy.
8. For More Please Refer Our PPT. Thank You.
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