1. (8 points) Gene x is a mouse gene that codes for protein x. You are intereste

Question

1. (8 points) Gene x is a mouse gene that codes for protein x. You are interested in studying the function of protein x. To do so, you will first need to clone gene x. Below is the sequence of gene x as it is found in mouse genomic DNA. The gene is flanked by several restriction enzyme recognition sites, as indicated. There are two plasmids that you can chose from for your cloning project, shown below. Each plasmid has several restriction enzyme recognition sites, as indicated. pUC17 has the amp gene (codes for a protein that confers ampicillin resistance). pUC18 has the tet gene (codes for a protein that confers tetracycline resistance). To work with, you also have E. coli cells and all of the enzymes and antibiotics that you could possibly need Fully describe the approach that you would use to clone gene x, including the enzymes, plasmids, and antibiotics that you would use at each step. Be specific. Use the steps of molecular cloning that we discussed in class as a guide. (Note: you won't be using blue/white screening for your selection, just talk about what antibiotic you would use for selection.) Ava T C 17 puC1

Explanation / Answer

Ans- the plasmid that can be used for the cloning of X gene is puc17 Because it has much more restriction sites available for the genes that's why it should be preferred. The antibiotics will be preferred ampicillin as the puc17 have ampR genes which is specific for ampicillin resistance. The enzyme will be used Ava 1 And Bam1 .

Steps the sequence will be cut with the help of enzymes and then it will be added to the plasmid with the help of ligase and after antubianti's treatment it will be left for overnight incubation.

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