Need assistance on that last question.. restriction sites, to make it easier to

Question

Need assistance on that last question..

restriction sites, to make it easier to insert foreign DNA. They also have other useful characteristics, such as resistance to ampicillin, which makes it possible to select bacteria that have been transformed, and not the far greater number of bacteria that were not transformed in the same process. Here is a diagram of one of the earlier plasmids to be engineered: pBR322 Notice all the restriction enzyme cut sites, which allow foreign DNA to be easily inserted. Hindlll Notice also the "amp" and "tet" sites. These allow the recipient bacteria to survive and reproduce Pstt Sall even when grown on media containing the ampicillin or tetracyclin, respectively tet amp Usually, when we transform bacteria, i.e. get them to take in our plasmid, most of the bacteria will not take in the plasmid. Before we used plasmids with antibiotic resistance genes in them, it was necessary screen many individual colonies to find the few bacteria that were transformed. But if the plasmid has an antibiotic resistance gene on it, the pBR322 bp ori transformed bacteria are the only ones that will grow on medium with the antibiotic in it. simplifying the work enormously. i' yau were to cur p8BR32 with Hind II, hat would huppen? If you were to cut pBR322 with EcoRV at 185 bp (base pairs i.e. nucleotides) and Nde I at 2295 bp, how many fragments would you get, and how big would they be, in base pairs? 147

Explanation / Answer

EcoRV = 185 bp

Nde I = 2295 bp

Both have a single recognition site. So, the double digestion would produce two bands.

1. 2110 bp

2. 2250 bp

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