PLEASE EXPLAIN You wish to make a recombinant DNA molecule that will contain one

Question

PLEASE EXPLAIN

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.

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

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 Answer AluI 1750 Answer EcoRI 2200 Answer NotI 3005 Answer 3005 Not I 2850 AbrII 2200 EcoRI 2055 XmaIII EcoRI BamHI Eco RI Spel Noti Xma III 255 375 225 320 230 Dral 120 Frog DNA EcoRI 510 BamHI 580 600 Not I 680 Spel pKABOO Xma III 720 3150bp AluI 1750 ori

Explanation / Answer

Answer:

The size of the recombinant plasmid is Vector size + insert size (here 55 base pair) - restriction or cloning site at vector which will be removed after restriction digestion.

So recombinant size will be =3150 + 55- 100 base = 3105 bases as you have to do the restriction digetion with SpeI and BamHI to both the vector and insert.

Restriction enzyme - Vector position- Recombinant position

Dral - 120 - 120

Alul - 1750 - 1805

EcoR1 - 2200 - 2200

Not I - 3005 - 3060

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