Bio 202 EXAM 3 QUESTIONS AND CORRECT DETAILED ANSWERS

Bio 202 EXAM 3 QUESTIONS AND CORRECT DETAILED ANSWERS How is recombinant DNA technology used to clone the human GH gene? 1st step! - ANSWER 1. use PCR to obtain the gene (Simply cutting the DNA with restriction enzymes would not be an effective way to isolate a specific gene. The genome would be cut into so many fragments, if you ran a gel it would be a smear! Which fragment is yours? You would need hybridization to isolate your fragment. Wouldn't have much DNA- not like the amount you can easily get from PCR.) The three main components of a cloning vector/plasmid - ANSWER 1 - origin of replication (digest vector and GH) 2- multiple cloning site 3- antibiotic resistance Transformation of recombinant DNA - ANSWER introduce recombinant DNA into bacteria so it replicates Steps of cloning - ANSWER 1. Isolated GH gene via PCR and engineered in EcoR1 sites with primers. 2. Ligated GH fragment into plasmid that is cut with EcoR1 at multiple cloning site. 3. Verified that we had some transformant clones with our GH gene through: blue white screen and digest or via PCR T/F cloning vectors can express a human vector - ANSWER False - an expression vector In the expression vector which promoter should be present to make the human GH protein in bacteria?? - ANSWER prokaryotic promoter Bacterial RNA polymerase will not recognize a eukaryotic promoter. Thus, we are making a hybrid: prokaryotic promoter + eukaryotic gene = recombinant DNA. How is GH cDNA different from GH genomic DNA? - ANSWER it contains only exons If a circular piece of DNA has three sites for a particular restriction enzyme, into how many fragments will that restriction enzyme cut the DNA? - ANSWER 3 If a linear piece of DNA has three sites for a particular restriction enzyme, into how many fragments will that restriction enzyme cut the DNA? - ANSWER 4 GMO - ANSWER genetically modified organsim - insect resistant - herbicide tolerant - drought tolerant - virus resistant Finding a naturally drought resistant plant and want to make GMO - ANSWER 1. Isolate DNA from the naturally occurring resistant plant 2. Amplify the gene that confers drought tolerance 3/4. Digest the naturally occurring drought resistant gene AND digest a plasmid 5. Create a recombinant DNA molecule 6. Transform Agrobacteria 7. Transform bean plant 8. Select for transformed bean plants Mendels second law - ANSWER law of independent assortment During gamete formation, the segregation of alleles at one locus is independent of the segregation of alleles at another locus. Dihybrid independent assortment Aa Bb - ANSWER 1/4 AB 1/4 Ab 1/4 aB 1/4 ab Dihybrid cross - ANSWER 9/16 both dominant 3/16 one dominant one recessive 3/16 one recessive, one dominant 1/16 both recessive The null hypothesis - ANSWER There is no significant difference between the observed and expected frequencies (i.e. any differences in observed values can be attributed to chance) A dihybrid was test-crosse