human insulin case studies
- Created by: olivia johnson
- Created on: 30-04-13 10:55
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- engineering case studies: human insulin
- pig insulin
- diadvantage: not identical to human insulin
- disadvantage: only small amount can be obtained from each pig
- small gene so difficult to locate.
- centrifugation used to obtain mRNA for gene. reverse transcriptase used to make a copy producing a single strand of DNA
- this is used as a template
- free nucleotides added to template of DNA to produce double stand called cDNA
- DNA polymerase
- plasmids cut open with restriction enzyme and mixed with cDNA insulin gene
- DNA ligase seals cDNA gene into cut plasmid
- recombinant plasmids taken up by bacteria
- some bacteria do not take up plasmid
- some bacterial plasmids sealed back together and not recombinant
- some bacteria do not take up plasmid
- recombinant plasmids taken up by bacteria
- if gene is present, bacteria will not be resistant to tetracycline but will be resistant to ampicillin
- replica plating-the process of growing bacteria on an agar plate then transfering a replica of that growth to other plates, usually containinng different growth promoters or inhibitors
- reverse transcriptase :retroviruses carry genetic material as RNA so when they infect a cell they cause reverse transcriptase to be synthesised in host cell.
- pig insulin
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