IGNOU LSE 3 2018 - What roles do the following play in cloning a DNA fragment?(a) Restr

Question

What roles do the following play in cloning a DNA fragment?
(a) Restriction enzymes.
(b) Vectors.
(c) DNA ligase.
(d) Electroporation.

23 Jan 2018

Answer :

Word Count : 472

(a) Restriction enzymes (restriction endonucleases) are proteins that cut DNA at (or close to) specific recognition sites (see the catalogs of manufacturers or the Restriction Enzyme Database). Two types of restriction enzymes exist that differ in the way they cut the target DNA:

Blunt end cutters. These enzymes cut both strand of the target DNA at the same spot creating blunt ends.

Sticky end cutters. These enzymes cut both strand of the target DNA at different spots creating 3'- or 5'-overhangs of 1 to 4 nucleotides (so-called sticky ends).

(b) A cloning vector is a small piece of DNA, taken from a virus, a plasmid, or the cell of a higher organism, that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes. The vector therefore contains features that allow for the

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