2023A3SEG23Wadidie

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University of Ottawa *

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3356

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Biology

Date

Dec 6, 2023

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docx

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2

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Assignement 1) Browsing the literature using Pubmed. Use the How to use Pubmed tutorial to answer the following questions: ( /2 Marks) a) Recent information: A scientific article on the inhibitory effect of theaflavin on the RNA polymerase of SARS virus family was published. Can you give me the Pubmed identifier (PMID) number of the article? Answer : PMID: 35566014 b) Past information: An article describing the sequencing and cloning of the SP6 RNA polymerase gene was published a long time ago. Can you give me the Pubmed identifier (PMID) number of this article? Answer : PMID: 3031606 c) Specific information: Can you give me the Pubmed identifier (PMID) number of the article published in 1995 discussing a T7 RNA polymerase mutant that can used a deoxyribonucleoside triphosphates as a substrate? Can you tell me the amino acid involved in this switch of specificity and its position in the protein? Answer : PMID: 7556104 The amino acid is the Tyrosine in Tyr639 d) Give a reference of a 2022, 2021 or 2020 scientific paper published by the professor listed for your section ( /0.5 Mark): Section E: John Pezacki Labana P, Dornan MH, Lafrenière M, Czarny TL, Brown ED, Pezacki JP, Boddy CN. Armeniaspirols inhibit the AAA+ proteases ClpXP and ClpYQ leading to cell division arrest in Gram-positive bacteria. Cell Chem Biol. 2021 Dec 16;28(12):1703-1715.e11. doi: 10.1016/j.chembiol.2021.07.001. Epub 2021 Jul 21. PMID: 34293284.
2) Subcloning Primer design is an essential skill for successful PCR amplification. Let say that you want to repeat this semester’s subcloning. Using the given coding sequence, choice of enzymes and vector, (a) design a forward and reverse primers to amplify the given coding sequence keeping in mind that you are planning to subclone it into the given vector ( /2 Marks). (b) Explain how the forward primer was (or wasn’t) modified to keep the reading frame of the amplicon in frame with the reading frame of the vector ( /2 Marks). Coding sequence of your given gene can be found on NCBI. Picture of the multiple cloning site of the different vectors are available on the virtual campus of the course ( Hypo vectors in Lab2 section).
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