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Introduction to Genetics

. A gene was ...

Question

. A gene was being ligated to the plasmid vector to prepare a recombinant DNA during
bacterial transformation. An exonuclease was added to the tube accidentally. How will it
affect the next step of the experiment?
What would happen if the restriction enzymes do not cut the DNA at specific recognition
sequences?

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Solution

Dear Student,

Here is the answer to you asked query-

The next step of the experiment will not be affected since the recombinant DNA molecule is circular and does not have any free ends. Therefore no substrate will be available for exonuclease enzyme to act and remove the nucleotides from the free ends of the given DNA. If the restriction enzyme does not cut the DNA at specific recogniton sequences than the desired gene can not be obtained as there will not sticky ends present and that specific character will also not be obtained.

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Similar questions

Q. Restriction enzymes that are used in the construction of recombinant DNA are endonucleases which cut the DNA at 'specific-recognition sequence'. What would be the disadvantage if they do not cut the DNA at specific-recognition sequence?

Q. Read the following statements and select the correct ones.

(i) The same kind of sticky ends are produced when DNA has been cut by different restriction enzymes.

(ii) Exonucleases make cuts at specific positions within the DNA.

(iii) Hind II was the first restriction endonuclease to be isolated.

(iv) A bacteriophage can replicate within bacterial cells by integrating its DNA with bacterial DNA.

(v) The presence of more than one recognition site for an enzyme within the vector complicates the gene cloning.

Q. Read the following statements and select the correct ones.
(i) Same kind of sticky ends are produced when a DNA is cut by different restriction enzymes.
(ii) Exonucleases make cuts at specific positions within the DNA.
(iii) Hind II was the first restriction endonuclease to be isolated.
(iv) A bacteriophage has the ability to replicate within bacterial cells by integrating its DNA with bacterial DNA.
(v) Presence of more than one recognition site for the same restriction enzyme within the vector complicates gene cloning.

Q. A piece of foreign DNA was inserted into a plasmid with an antibiotic resistance gene and lactose Z gene. The plasmid DNA was cut with a restriction enzyme, which splits the lactose Z gene and opens the circle. The foreign DNA was next inserted into the open restriction site of the plasmid. When the recombinant plasmid was introduced into bacterial cells and grown in the presence of antibiotic, some of the colonies turned blue in the presence of gene. The blue colonies contained

Q. From the below list, which of the following is the most logical sequence of steps for splicing foreign DNA into a plasmid and inserting the plasmid into a bacterium?
(i) Transformbacteria with recombinant DNA molecule.
(ii) Cut the plasmid DNA using restriction enzymes.
(iii) Extract plasmid DNA from bacterial cells.
(iv) Hydrogen-bond the plasmid DNA to nonplasmid DNA fragments.
(v) Use ligase to seal plasmid DNA to nonplasmid DNA.

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