Updated: 13 September, 2006
 
  Bio 184 Practical 1 Lab 1

Restriction Enzymes

 

 


Definition

A restriction enzyme (or restriction endonuclease) is an enzyme that cuts double-stranded DNA. The enzyme makes two incisions, one through each of the phosphate backbones of the double helix without damaging the bases. The chemical bonds that the enzymes cleave can be reformed by other enzymes known as ligases, so that restriction fragments carved from different chromosomes or genes can be spliced together, provided their ends are complementary (more below). Many of the procedures of molecular biology and genetic engineering rely on restriction enzymes. The term restriction comes from the fact that these enzymes were discovered in E. coli strains that appeared to be restricting the infection by certain bacteriophages. Restriction enzymes therefore are believed to be a mechanism evolved by bacteria to resist viral attack and to help in the removal of viral sequences.

 

Ligases


Definition

In biochemistry, a ligase (from the Latin verb lig?re — "to bind" or "to glue together") is an enzyme that can catalyse the joining of two large molecules by forming a new chemical bond, usually with accompanying hydrolysis of a small chemical group pendant to one of the larger molecules. Generally ligase catalyses the following reation:

Ab + C → A–C + b

or sometimes

Ab + cD → A–D + b + c

where the lower case letters signify the small, pendant groups.


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