Question

other enzymes used in cloning_?​

Answer 1

Answer:

$\underline \mathscr \purple{Question \: : - }$

other enzymes used in cloning_?

$\underline \mathscr \purple{Answer \: : - }$

Alkaline phosphatase: Ligation requires the presence of a 5'-phosphate group. If some of the fragments are treated with alkaline phosphatese to remove their phosphate groups then these cannot ligate within themselves and are forced to ligate with other fragments containing 5'phosphate group. Hence, this is a useful strategy to prevent self-ligation which would otherwise lead to wasteful ligation of fragments treated with restriction enzymes. An insert is ligated to the vector in generating rDNA as the vector is prevented from self-ligation by treating it with alkaline phosphatase. Alkaline phosphatase used for this purpose is purified from bacteria or calf intestines.

Answer 2

The activities of the cloning enzymes consist of ligases, kinases, phosphatases, and RecA Protein. DNA ligase, for example is used in joining DNA molecules together. Within the cell, the double ends of the strands are attached covalently.

A common method uses two types of enzymes: restriction enzymes and DNA ligase. A restriction enzyme is a DNA-cutting enzyme that recognizes a specific target sequence and cuts DNA into two pieces at or near that site. Many restriction enzymes produce cut ends with short, single-stranded overhangs.

Zinc finger nucleases are the most commonly used artificial restriction enzymes and are generally used in genetic engineering applications, but can also be used for more standard gene cloning applications.