EcoRV crystal structure complexed with double-stranded DNA

EcoRV (pronounced "eco R five") is a nuclease enzyme isolated from certain strains of E. coli, A nuclease is an enzyme capable of cleaving the phosphodiester bonds between the nucleotide subunits of nucleic acids. ... Binomial name Escherichia coli T. Escherich, 1885 Escherichia coli (usually abbreviated to E. coli) is one of the main species of bacteria that live in the lower intestines of warm-blooded animals (including birds and mammals) and are necessary for the proper digestion of food. ...

In molecular biology, it is a commonly used restriction enzyme. It creates blunt ends. The enzyme recognizes DNA the 6-base sequence 5'-GAT|ATC-3' and makes a cut at the vertical line. The complementary sequence is then 3'-CTA|TAG-5'. The ends are blunt and can be ligated into a blunt cloning site easily but with lower efficiency then sticky ends. Molecular biology is the study of biology at a molecular level. ... 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. ... In biology, sticky end and blunt end are the two possible configurations resulting from the breaking of double-stranded DNA. DNA exhibits a stabilizing interaction between complementary base pairs, providing specificity to the pairing of two strands of DNA. If two complementary strands of DNA are of equal length, then...

Structure

Structure of EcoRV (green) in a complex with its substrate DNA. Created from PDB 1RVA.

EcoRV has been cocrystallized with the sequence it normally cuts. This crystal was used to solve the structure [1] of the complex.

Mode of action

Like EcoRI, EcoRV forms a homodimer in solution before binding and acting on its recognition sequence. In molecular biology, EcoRI (pronounced Eco R One) is a commonly used restriction enzyme. ... Sucrose, or common table sugar, is composed of glucose and fructose. ... The recognition sequence of any DNA-binding protein motif that exhibits binding specificity, refers to the DNA sequence (or subset thereof), to which the domain is specific. ...

Uses

EcoRV is often used to cut open a plasmid vector to insert a gene-of-interest during gene cloning. The enzyme is supplied by many manufacturers and requires bovine serum albumin to work properly. Figure 1: Schematic drawing of a bacterium with plasmids enclosed. ... This stylistic schematic diagram shows a gene in relation to the double helix structure of DNA and to a chromosome (right). ... Cloning is the process of creating an identical copy of an original. ... Bovine serum albumin (BSA) can be used as a diluent or a blocking agent in numerous applications including ELISAs (Enzyme-Linked Immunosorbent Assay), blots and immunohistochemistry. ...

See also

• EcoRI, another nuclease enzyme from 'E. coli.
• FokI, a nuclease enzyme from Flavobacterium okeanokoites

In molecular biology, EcoRI (pronounced Eco R One) is a commonly used restriction enzyme. ...

References

• Bitinaite, J., D. A. Wah, Aggarwal, A. K., Schildkraut, I. (1998). "FokI dimerization is required for DNA cleavage." Proc Natl Acad Sci U S A 95(18): 10570-5.