Bio1_19_DNAtech - 1 2 3 4 5 6 7 8 Self-Quiz 1. What is the...

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1 Self-Quiz 1. What is the function of each of the 3 main classes of RNA? 2. What does RNA polymerase do? 3. Describe transcription base pairing. 4. What are removed during transcript processing? 5. Describe the 3 steps in translation. 6. What are the possible effects of gene mutations? Overview: DNA – DNA cloning – Biotechnology products – Gene therapy – Genomics DNA Cloning • Gene cloning – Produces genetically identical copies of DNA – Gene therapy: cloned genes in humans – Transgenic organisms: cloned genes in other species • Procedures to clone DNA – Recombinant DNA technology (rDNA) – Polymerase chain reaction (PCR) Recombinant DNA • Plasmid – Small circle of DNA outside the chromosome • Cloning vector – Recombinant plasmid put into bacteria, yeast, etc. – Plasmid replicates with cells Recombinant DNA (rDNA) • Restriction enzymes – Cuts DNA at specific sequence – Sequence same on both strands – “Sticky” ends • DNA ligase – Seals nicks Assignment 1 • Topic: A restriction site is typically a “palindromic” recognition site – Both strands with same sequence in 5’ b 3’ 5’ – GAATTC – 3’ 3’ – CTTAAG – 5’ • Write the complement & box the palindromic site for the following: 5’ – TGAATCTAGACGGTT – 3’ 3’ – ACTTAGATCTGCCAA – 5’ Bacterial Expression of Gene • Plasmid must contain regulatory regions • Using eukaryotic genes: mRNA b cDNA – Complimentary DNA – Reverse transcriptase •Enzyme from viruses •Makes DNA from RNA – Compliment of cDNA •Double strand •DNA polymerase – No introns in cDNA Assignment 2 • Think, Pair, Share 1 2 3 4 5 6 7 8
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2 • Topic: cDNA is made from mRNA using reverse transcriptase: mRNA: 5’ – CAUGAU – 3’ cDNA: 3’ – GTACTA – 5’ • Determine the cDNA strand for each of the following: – 5’ – UUACCG – 3’ – 5’ – CUAAUG – 3’ Amplifying DNA • Polymerase chain reaction (PCR) – Many copies of target DNA – Primers bracket desired sequence – Heat-resistant DNA polymerase • Primers – Get each copy started
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This note was uploaded on 01/12/2012 for the course BIOLOGY 1 taught by Professor Teresafinnern during the Fall '11 term at Riverside Community College.

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Bio1_19_DNAtech - 1 2 3 4 5 6 7 8 Self-Quiz 1. What is the...

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