(a) In pBR322, foreign DNA has to be introduced in tetRregion. From the restriction enzymes given below, which one should be used and why: PvuI, EcoRI, BamHI (b) Give reasons, why the other two enzymes cannot be used?
class 12th CBSE Biology sample paper
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(a)The restriction enzymes that should be used are Bam HI , as restriction site for this enzyme is present in tetR region.
Bam HI is a type II restriction endonuclease, having the capacity for recognizing short sequences of DNA and specifically cleaving them at a target site.
(b) PvuI will not be used, as restriction site for this enzyme is present in ampR region (not in tetR)
EcoRI is a restriction endonuclease enzyme isolated from species E. coli and also not used.The Eco part of the enzyme's name originates from the species from which it was isolated,
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(a)The restriction enzymes that should be used are Bam HI , as restriction site for this enzyme is present in tetR region.
BamHI( from BAcillus amyloli) is a type II restriction enzyme that derived from B. amyloliquefaciens , having the capacity for recognizing short sequences of DNA and specifically cleaving them at a target site. In order for restriction endonuclease to bind to their target sequence, they have to first bound to a nonspecific DNA.
(b) PvuI will not be used, as restriction site for this enzyme is present in ampR region (not in tetR)PvuI has a High Fidelity version PvuI-HF® (NEB #R3150). High Fidelity (HF) Restriction Enzymes have 100% activity in CutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing.
EcoRI is a restriction endonuclease enzyme isolated from species E. coli and also not used.The Eco part of the enzyme's name originates from the species from which it was isolated,EcoRI is a restriction endonuclease enzyme isolated from species E. coli. The Eco part of the enzyme's name originates from the species from which it was isolated, while the R represents the particular strain, in this case RY13.
-Ekansh Nimbalkar
BamHI( from BAcillus amyloli) is a type II restriction enzyme that derived from B. amyloliquefaciens , having the capacity for recognizing short sequences of DNA and specifically cleaving them at a target site. In order for restriction endonuclease to bind to their target sequence, they have to first bound to a nonspecific DNA.
(b) PvuI will not be used, as restriction site for this enzyme is present in ampR region (not in tetR)PvuI has a High Fidelity version PvuI-HF® (NEB #R3150). High Fidelity (HF) Restriction Enzymes have 100% activity in CutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing.
EcoRI is a restriction endonuclease enzyme isolated from species E. coli and also not used.The Eco part of the enzyme's name originates from the species from which it was isolated,EcoRI is a restriction endonuclease enzyme isolated from species E. coli. The Eco part of the enzyme's name originates from the species from which it was isolated, while the R represents the particular strain, in this case RY13.
-Ekansh Nimbalkar
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