| Term | Definition |
|---|
| biotechnology | the use of microorganisms, cells, or cell components to make a product |
| recombinant DNA technology | a type of biotechnology; insertion of genes to produce desired proteins |
| bacterial transformation | inserting exogenous DNA into bacteria so it transcribes and translates desired protein |
| plasmid | secondary source of DNA outside of chromosome that carry special genes that give it some advantage over nature (example resistance) |
| vector | mode of transportation |
| restriction enzyme | act on DNA by cutting it; act on plasmid by opening it; must use same restriction enzyme for plasmid and gene of interest |
| bacterial transformation Step 1 | vector such as plasmid is isolated |
| bacterial transformation Step 2 | DNA is cleaved by enzyme into fragments |
| bacterial transformation Step 3 | gene is insterted into plasmid - forms recombinant DNA |
| bacterial transformation Step 4 | plasmid is taken up by bacterium - forms recombinant bacterium |
| bacterial transformation Step 5 | cells with gene of interest are cloned |
| bacterial transformation Step 6a | copies of gene are harvested |
| bacterial transformation Step 6b | cells make a protein product |
| bacterial transformation Step 7 | copies of protein are harvested |
| bacterial transformation gene harvest example | gene for pest resistance inserted into plants; gene alters bacteria for cleaning up toxic waste |
| bacterial transformation protein harvest example | enzymes used to prepare clothing for manufacture; human growth hormone treating stunted growth |
| how restriction enzymes work | cut DNA at recognition site; produce DNA fragment w/ 2 sticky ends; 2 fragments cut w/ same restriction enzymes join by base pairing; joined fragments form linear or circular molecule; DNA ligase unites backbones of DNA fragments, producing a recombinant DNA molecule |
| Herb Boyer and Robert Swanson | form Genentech - produce human insulin in 1976 |
| Kary Mullis | PCR in 1983 (won Nobel prize in 1993) |
| polymerase chain reaction (PCR) | based on premise of how DNA replicates in body; makes multiple copies of a piece of DNA |
| PCR machine | cyclical temperature machine (heating and cooling) |
| PCR Step 1 | incubate target DNA at 94C for 1 min to separate strands - hydrogen bonds denature at high temperatures |
| PCR Step 2 | add primers (small stretches of DNA that accelerate the process), nucleotides, and DNA polymerase |
| PCR Step 3 | incubate at 60C for 1 min - temp causes primers to attach |
| PCR Step 4 | incubate at 72C for 1 min, 2 copies of DNA are formed - increase in temp causes DNA polymerase to activate |
| PCR Step 5 | repeat heat and cooling cycle to make 2 more copies of target DNA |
| PCR applications | clone DNA for recombination; amplify DNA to detectable levels |
Combine with other sets
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