You wish to make a recombinant DNA molecule that will contain one piece of pKABO

Question

You wish to make a recombinant DNA molecule that will contain one piece of pKABOO vector DNA and one piece of frog DNA so that you can clone a segment of frog DNA. You want cells containing your recombinant plasmid to be ampr and tets and you want to use enzymes that cut within the insertional marker gene. (Note that tets means that there is no functional tetr gene in the plasmid.) Be sure that your plasmid has the ability to replicate autonomously in a bacterial cell. You do not have to include the entire frog DNA given below in your recombinant plasmid.

1a) What restriction enzymes would be used to clone a segment of frog DNA?

A. BamHI and Hindlll

B. Hindlll and Pstl

C. Spel and Hindll

D, BamHI and PstI

E. BamHl and Spel

F. Spel and Pstl

1b) What is the size of the recombinant plasmid (in bp)?

1c) The recombinant plasmid when transformed into E. coli confers resistance to which of the following antibiotics:

A. ampicillin and beta-galactocillin

B. beta-galactocillin only

C. ampicillin only

D. tetracycline and beta-galactocillin

E. ampicillin and tetracycline

F. tetracycline only

1e) The recombinant plasmid map has many restriction sites. Determine the base pair position of each of the following sites in the recombinant plasmid map:

Restriction Enzyme Vector Position Recombinant Position NotI 75 Answer AluI 1050 Answer BamHI 1525 Answer PstI 2150 Answer BamHI KpnI SpeI EcoRI HindIII PstI 350 400 200 300 700 NotI 75 2150 PstI Frog DNA 550 600 BamHI SpeI HindIII 750 PstI ClaI 1900 KpnI pKABOO 2500bp 900 915 1525 BamHI AluI 1050 1200 EcoRI ori

Explanation / Answer

1 a) C

1 b) 2850bp

1 c) C

1 e) NotI 75 75

AluI 1050 1400

BamHI 1525 1525

PstI 2150 2500

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