ECORV (pronounced "eco R five") is a type II restriction endonuclease isolated from certain strains of Escherichia coli . It has the alternative name Eco32I.
In molecular biology , it is a commonly used restriction enzyme . It creates blunt ends . The enzyme recognizes the palindromic 6-base DNA sequence 5'-GATATC-3' and makes a cut at the vertical line. The complementary sequence is then 3'-CTATAG-5'. The ends are blunt and can be ligated into a blunt cloning site easily but with lower efficiency than sticky ends.
* 1 Structure * 2 Mode of action * 3 Uses * 4 See also * 5 References
The structure of this enzyme, and several mutants, in complex with the DNA sequence which it cuts has been solved by X-ray crystallography .
The core of the enzyme consists of a five-stranded mixed β-sheet
flanked by α-helices. The core is conserved in all other type II
restriction endonucleases. It also has an N-terminal dimerization
subdomain formed by a short α-helix, a two-stranded antiparallel
-sheet, and a long α-helix. This subdomain is found only in
MODE OF ACTION
Binding of the enzyme induces a conformational change in the DNA, bending it by about 50°. DNA bending results in the unstacking of the bases, widening of the minor groove, and compression of the major groove. This brings the phosphodiester linkage to be broken closer to the active site of the enzyme, where it can be cleaved. Cleavage occurs within the recognition sequence, and does not require ATP hydrolysis.
* ^ A B C D Pingoud A, Jeltsch A (2001). "Structure and function of type II restriction endonucleases" . Nucleic Acids Research. 29 (18): 3705–3727. PMC 55916 . PMID 11557805 . doi :10.1093/nar/29.18.3705 . * ^ Bitinaite J, Wah DA, Aggarwal AK, Schildkraut I (1998). "FokI dimerization is required for DNA cleavage" . Proc Natl Acad Sci USA. 95 (18): 10570–10575. PMC 27935 . PMID 9724744 . doi :10.1073/pnas.95.18.10570 . * ^ Zahran, M., Daidone, I., Smith, J. C., ;background:none transparent;border:none;-moz-box-shadow:none;-webkit-box-shadow:none;box-shadow:none;">v
* t * e
Hydrolase : esterases (EC 3.1)
3.1.1 : Carboxylic ester hydrolases
* Bile salt-dependent * Gastric /Lingual * Pancreatic * Lysosomal * Hormone-sensitive * Endothelial * Hepatic * Lipoprotein * Monoacylglycerol * Diacylglycerol
* A1 * A2 * B
3.1.2 : THIOESTERASE
3.1.3 : PHOSPHATASE
* IMPA1 , IMPA2 , IMPA3
* C * D
3.1.6 : SULFATASE
Nuclease (includes deoxyribonuclease and ribonuclease )
3.1.11 -16: Exonuclease
3.1.21 -31: Endonuclease
* 1 * 2A * 2B * 2C
* RNase A
* 1 * 2 * 3 * 4/5
EITHER DEOXY- OR RIBO-
* v * t * e
* EC1 OXIDOREDUCTASES (list ) * EC2 TRANSFERASES (list ) * EC3 HYDROLASES (list ) * EC4 LYASES (list ) * EC5 ISOMERASES (list ) * EC6 LIGASES (list )
* Molecular and Cellular Biology portal