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प्रश्न
A plasmid without a selectable marker was chosen as vector for cloning a gene. How does this affect the experiment?
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उत्तर
In a gene, cloning experiment, first a recombinant DNA molecule is constructed, where the gene of interest is ligated to the vector (the step would not be affected) and introduced inside the host cell (transformation). Since, not all the cells get transformed with the recombinant/plasmid DNA, in the absence of a selectable marker, it will be difficult to distinguish between transformants and non-transformants, because role of the selectable marker is in the selection of transformants.
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संबंधित प्रश्न
State how has Agrobacterium tumifaciens been made a useful cloning vector to transfer DNA to plant cells.
Draw a schematic sketch of pBR 322 plasmid and label the following in it:
(a) Any two restriction sites.
(b) Ori and rop genes.
(c) An antibiotic resistant gene.
Can you think and answer how a reporter enzyme can be used to monitor the transformation of host cells by foreign DNA in addition to a selectable marker?
Describe briefly the following:
Origin of replication
Why is the coding sequence of an enzyme β-galactosidase a preferred selectable marker in comparison to the ones named above?
Write the role of ‘restriction sites’ in the cloning vector pBR322.
What is ‘Ori’ ? State its importance during cloning of a vector.
Answer the following question.
Expand ‘YAC’ and mention what it was used for.
How does the β-galactosidase coding sequence act as a selectable marker? Why is it a preferred selectable marker to antibiotic resistance genes? Explain.
The genetic material of virus includes:
Which of the following is correctly matched?
Which of the following statement is not true for a clone?
A suitable vector for gene cloning in higher organism is ______.
Plasmid pBR322 has a PstI restriction enzyme site within gene ampR that confers ampicillin resistance. If this enzyme is used for inserting a gene for β-galactoside production and the recombinant plasmid is inserted in an E.coli strain.
What does ‘competent’ refer to in competent cells used in transformation experiments?
Name the regions marked A, B and C.
For selection of recombinants, insertional inactivation of antibiotic marker has been superceded by insertional inactivation of a marker gene coding for a chromogenic substrate. Give reasons.
