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Question: answer the following questions could be more than one...

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Answer the following questions ( could be more than one answer)

Question 15 Commonly used plasmids are: Reconstructed using parts of original plasmids occur in bacteria. ■ carry essential genes for the bacteria. Replicate rarelin transformed cells Produce antibiotics found in nature. ■ .
Question 18 Applications and advantages of phagemid vectors: Cloning and storing genes Expressing genes of interest. Sequencing genes from the library Target gene may be inserted into E. coli genome and other genome. > Moving to th
Question 19 Describe the roles of M13 genes and their proteins. Genes 1, 4 and 11 produces proteins required for assembling the the phage. a Genes 3.6,7,8 and 9 produces capsid proteins Gene 2 nicks the circular phage ssDNA to make it double stranded by DNA polymerase Gene 5 produces single strand binding proteins and gene 1o protein makes M13 SSDNA dircular. ■ Protein five bound single stranded circular M13 moved to the E. coli membranne and is released out coated will proteins 3, 6, 7, 8 and 9
Question 21 What is a phagemid and how does it work? a it is a recombinant DNA with afilamentous phage genome and a plasmid genome. 目it carries two origins of replication: eg., or fi and on ColE1. 0 May carry a gene for selection: e.g. Ampr gene. O May carry a marker gene; e.g, lacZ gene with promoter and regulator. ■ Mary contain MCS, as is pUC18.
Question 22 o toi n A vector DNA. D A cloning site in the vector matching with the ends of a gene to be cloned. A host organism capable to hold the recombined vector. D A marker gene present in the recombinant vector G A functional gene of interest.
Question 24 identify. which of the following is true for the bacteriophage lambda. D It has two major body parts, the head and the tail. The genome is approximately 49 thousand bases long. a It has a linear dsDNA with single stranded 12 bases long cos terminals at both a The single stranded lambda DNA cos terminals appear to be as follows: (a) GGGCGGCGACCT and (b)CCCGCCGCTGGA a it can accept 7-10 kb long foreign DNA replacing its non-essential b2 region.
Question 26 Which of the following statements is/are incorrect? The meaning of virus is poison. Bacteriophages refer to virus for bacteria. Bacteriophage means food for bacteria. Bacteria eaters refer to bacteriophage Moving to the next question prevents changes t o thiem
Question 29 For developing a recombinant lambda DNA with a gene of interest and expressing the gene in a host you have to take the following steps: D Remove the b2 region from lambda DNA using a restriction enzyme. Digest the gene of interest, outside its stop and start codons, with the same restriction enzyme. Recombine the lambda DNA digest with the digested gene of interest carrying complementary overhangs with ligase. Form lambda particles by adding head proteins, tail proteins and packaging enzynes to the recombined lambda DNA. Add the packaged lambda particles to Agrobacetrium tumefaciens and incubate them in culture. Add the packaged lambda particles to laboratory strains of Escherichia coli and incubate them in culture. (a
Question 32 The amino acids coded by our genes differ by the following Polarity. Acidity Basicity Aroma. Hydrophobicity. A Moving to the nout
Question 38 What is an M13 phage and what are its specialty? They are rod shaped Escherichia coli bacteriophage. ■ its genome is made of a single stranded circular DNA. 0 Its DNA is 6400 nucloetides long coding for eleven proteins. After infecting a bacterial cell it starts replicating in 10 minutes producing 1,000 phages in an ho □ soon its kills the host Escherichia coli and gets out to the environment.
uestion 40 You have a gene with BamH cut sites out side its start and the stop codons. You can use the follovwing procedures to prepare a clone of the gene in pBRszz and select th ed Ecof Digest both the gene and the plasmid pR322 with BamH1 Ligate the fragments using T4 DNA ligase. O Transform E coli with the reconstructed plasmid O Select the cells tarnsformed cells grow in L8-agar-ampicilin plates O Select the cells transformed cells grow in LB-agar-tetracycline plates
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