HC70A quiz 2 - Name Marissa Lara Individuals in Group(if...

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Name: Marissa Lara Individuals in Group (if any): HC70A/SAS70A WINTER 2016 PROFESSORS BOB GOLDBERG & JOHN HARADA QUIZ #2 Due Tuesday, January 19 at the Beginning of Class Follow Instructions For Quiz #1 Note: Be Prepared to Answer Quiz Questions in Class! QUESTION ONE – 10,000 points The invention of cDNA cloning in 1976 by scientists working independently at UCLA and Harvard started a second genetic engineering revolution as it provided a “pathway” towards isolating specific genes, studying their functions, and engineering bacteria to synthesize medically relevant human proteins. It also lead, indirectly, to the start of a multi-billion dollar biotechnology industry based on drugs made using recombinant cDNAs that have improved the health of millions of people around the globe! A. OUTLINE the specific role each component of the genetic engineer’s toolbox performs in generating recombinant cDNA molecules. 1. Reverse transcriptase: 1. An enzyme used to catalyze the synthesis of single stranded DNA from the mRNA. 2. S1 Nuclease: 1. Separates, recognizes the single strand into 2 strands 2. Opens the “loop” generated from the synthesis of the cDNA, and through the opening or cutting of the loop, cloning is possible. 3. Terminal Transferase: Produces cohesive ends, creating overhangs, by catalyzing the addition of deoxynucleotides to the three prime end of DNA molecules. 4. DNA polymerase: An enzyme found in DNA replication, which from the loop can produce a complementary sequence for the cDNA. 5. mRNA: 1. cDNA’s are synthesized from mRNA using the enzyme reverse transcriptase. 2. mRNA is the molecule which serves as the template from which cDNA is synthesized B. OUTLINE conceptually the difference between a library of cDNA clones and a library of genomic clones.
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