7- Name three features that a cloning vector for use in bacteria must contain. 1- 2- 3- 8- Briefly state why heat-stable DNA polymerase from a thermophilic bacterium is used in PCR instead of DNA polymerase from E. coli or humans. 9- How many DNA copies will be produced after 5 cycles of PCR? 10- Genes are evenly distributed throughout the human genome. (true, false) 11- Dark staining chromosome bands are correlated with protein synthesis. (true, false) 12- Stem cells with the maximum flexibility for expression are ____ (embryonic, tissue-specific) in nature. 13- Give two reasons why the drug companies who produce insulin wanted to clone the human insulin gene when they already had a reasonably cheap source of the hormone (pig insulin). 1- 2- 14- Which of the following reactions are known to be carried out by a ribozyme? a) DNA synthesis; b) transcription; c) RNA splicing; d) protein hydrolysis 15- Which of the following pairs of codons might you expect to be read by the same tRNA as a result of wobble? a) CUU & UUU; b) GAU & GAA; c) CAC & CAU; d) AAU & AGU; e) CCU & GCU
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Use this image of a generic cloning vector to answer questions 21-23. HindIII SphI PstI DNA sequencing SalI AccI HincII primer site XbaI 21 BamHI Multiple Smal cloning site Xmal Kpnl BanII Ampicillin lacZ SstI resistance gene EcoRI 23 gene (ampR) DNA sequencing primer site Origin of 22 replication (ori) 21) What is the purpose of a multiple cloning site? a. allows the vector to replicate independent of the host's genome b. contains known sequences to allow the vector to be sequenced c. the region in which a DNA fragment is cut and pasted d. a selectable marker to identify cells transformed by the vector 22) What is the purpose of the origin of replication? a. allows the vector to replicate independent of the host's genome b. contains known sequences to allow the vector to be sequenced c. the region in which a DNA fragment is cut and pasted d. a selectable marker to identify cells transformed by the vector 23) What is the purpose of the ampicillin resistance gene? a. allows the vector to replicate independent of the host's genome b. contains known sequences to allow the vector to be sequenced c. the region in which a DNA fragment is cut and pasted d. a selectable marker to identify cells transformed by the vector
Sri K.
a. Most bacterial cells are naturally competent, so DNA just needs to be mixed with the cells. b. Many bacterial cells can be transformed after being made competent by treatment with cold CaCl followed by a brief heat shock at 43°C. c. Many bacterial cells can be transformed by applying high voltage to a mixture of cells and DNA for a sustained period of time. d. Transformation can occur if DNA is mixed with bacterial cells that have been heat shocked. e. None of these choices are correct.
Keemin L.
Describe the process of molecular cloning. a. The foreign DNA and plasmid are cut with the same restriction enzyme and DNA is inserted within the lacZ gene, whose product metabolizes lactose. The foreign DNA and vector are allowed to anneal. The vector is transferred to a bacterial host that is ampicillin sensitive and those with a disrupted lacZ gene show inability to metabolize X-gal. b. The foreign DNA and plasmid are denatured using high heat, and DNA is inserted within the lacZ gene, whose product metabolizes glucose. The foreign DNA and vector are allowed to anneal. The vector is transferred to a bacterial host that is ampicillin sensitive and disrupted lacZ gene will metabolize X-gal c. The foreign DNA and plasmid are cut with the same restriction enzyme and DNA is inserted randomly in the plasmid. The foreign DNA and vector are allowed to anneal. The vector is transferred to a bacterial host that is ampicillin sensitive and the disrupted lacZ gene shows inability to synthesize X-gal. d. The foreign DNA and plasmid are cut with the same restriction enzyme and DNA is inserted within the lacZ gene, whose product metabolizes lactose. The foreign DNA and vector are allowed to anneal. The vector is transformed into a viral host that is ampicillin sensitive and the disrupted lacZ gene show inability to synthesize X-gal.
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