Lecture 5 Genetic Engineering Flashcards
define GMO
genetically modified organism- an organism with an altered genome. allows for expression of eukaqryotic genes in prokaryotes i.e insulin.
achieved by cloning th gene in mRNA
explain the process of expressing a recombinant mammalian protein
- identification of gene
- generating cDNA (complementary DNA) from the mRNA to create a vector
- selection of a suitable system that can express the vector
- methods to scale up the gene
what is the purpose of plasmids in genetic engineering
plasmids can uptake the gene of expression, the cells with the positive clone can then be selected and expanded. cells and components can then be isolated and purified to gather the secreted/intracellular components
what issues may arise from expressing mammalian genes in bacteria?
the synthesised proteins are subject to a lot of problems, i.e
- degradation by intracellular proteases
-toxicity to the prokaryotic host
- formation of inclusion bodies (by products) when the bacteria are induced to overexpress recombinant protein
what does RNA polymerase, antibiotic resistance, and origin of replication do for the gene and plasmid
RNA poly binds at the promoter and synthesises the gene
Antibiotic resistance allows for selection in host
Ori allows the plasmid to replicate
write a note on genetically engineered somatotropin
somatotropin= growth hormone
-commonly used in dairy industry (recombinant bovine somatotropin) to stimulate milk production in cows
what are some human recombinants used today!
human growth hormone (rHGH): humatrope from Lilly that replaces cadaver harvested HGH (pituatory glands)
human insulin: humulin from Lilly and novolin from novo nordisk now replace bovine and porcine insulin for human use
FSH (follicle stim hormone) used as a gonadotropin replaces isolated FSH from post menopausal female urine
Factor V111: is now replaced from previously blood harvested factor viii
