In $p B R 322$ , foreign DNA has to be introduced in $t e t^{R}$ region. From the restriction enzymes given, which one should be used and why: Pvul, EcoRI, BamHI.

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Solution

Restriction enzyme BamHI should be used because the restriction site for this enzyme is present within the gene coding for tetracycline resistance, using any other enzyme would not lead to insertional inactivation of tetracycline resistance coding gene, which is later used for transformant and recombinant identification.

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Similar questions

Q. In a pBR 322, a foreign DNA segment has to be introduced in the

t e t^{R}

region. From the restriction enzymes,

B a m H I

and

E c o R I

, which one would you select for use? Give reason. Also mention why not the other one.

Q. Assertion :E.coli having pBR322 with DNA insert at BamHI site cannot grow in medium containing tetracycline. Reason: Recognition site for BamHI is present in

t e t^{R}

region of pBR322.

Q. Consider the linear double-stranded DNA shown below. The restriction enzyme sites and the lengths demarcated are shown. This DNA is completely digested with both EcoRI and BamHI restriction enzymes. If the product is analyzed by gel electrophoresis, how many distinct bands would be observed?

Q. Assertion :Recognition site should be preferably single and responsive to commonly used restriction enzymes. Reason: In

p B R 322

alien

D N A

is ligated generally in the area of

B a m - H I

site of tetracycline resistance gene.

Consider the linear double stranded DND shown below. The restriction enzyme sites and the lengths demarcated are shown. This DNA is completely digested with both EcoRI and BamHI restriction enzymes. If the Product is analyzed by gel electrophoresis, how many distinct bands would be observed?

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In pBR322, foreign DNA has to be introduced in tetR region. From the restriction enzymes given, which one should be used and why: Pvul, EcoRI, BamHI.

In $p B R 322$ , foreign DNA has to be introduced in $t e t^{R}$ region. From the restriction enzymes given, which one should be used and why: Pvul, EcoRI, BamHI.