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.

1) Restriction enzymes that would be used to clone a segment of frog DNA:

A) EcoRI and BamHI,

B) EcoRI only

C) EcoRI and SpeI

D) EcoRI and NotI

E) EcoRI and XmaIII

F) BamHI and SpeI

G) BamHI and NotI

H) BamHI and XmaIII

I) SpeI and NotI

J) SpeI and XmaIII

K) NotI and XmaIII

2) The size of the recombinant plasmid is _____ bp.

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

a) ampicillin only

b) tetracycline only

c) beta-galactocillin only

d) ampicillin and tetracycline

e) ampicillin and beta-galactocillin

f) tetracycline and beta-galactocillin

4) 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 DraI 120 _____ AluI 1750 _____ EcoRI 2200 _____ NotI 3005 _____ 3005 NotI 2850 Abr II 2200 EcoRI 2055 Xma III EcoRI BamHI EcoRI Spel Not I Xma III 255 375 225 320 230 DraI 120 Frog DNA. 510 EcoRI 580 BamHI 600 NotI 680 Spel pKABOO Xuma III 720 3150bp AluI 1750 ori

Explanation / Answer

1: D: Restriction enzymes that would be used to clone a segment of frog DNA are EcoRI and NotI
2: The size of the recombinant plasmid is:
3005bp -510 bp=2495 bp
Size of original plasmid= 3150 bp
Size of plasmid after removal of 2495 bp= 3150 bp-2495 bp= 655 bp
Size of recombinant plasmid= Size of plasmid + insert= 655 + 225 + 320= 1200bp
3: The recombinant plasmid when transformed into E. coli confers resistance to which of the following antibiotics:
a) ampicillin only

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