NotI
| notI ( Nocardia otitidiscaviarum ) | ||
|---|---|---|
| Mass / length primary structure | 383 amino acids | |
| Secondary to quaternary structure | Homodimer | |
| Identifier | ||
| Gene name (s) | notI (Rebase) | |
| External IDs | ||
| Enzyme classification | ||
| EC, category | 3.1.21.4 , restriction enzyme | |
| Response type | hydrolysis | |
| Substrate | DNA | |
| Products | Double stranded DNA fragments with terminal 5 'phosphate groups | |
The enzyme Not I is a Type II restriction enzyme , an endonuclease that cuts DNA at a specific site. The enzyme was isolated from the bacterium Nocardia otitidis-cavarium . It has a sequence-specific recognition site on the DNA to which it can specifically bind and cut the DNA at this point. The corresponding gene was sequenced and cloned.
| Recognition sequence | Restriction cut |
|---|---|
5'-GCGGCCGC-3' 3'-CGCCGGCG-5' |
5'-GC GGCCGC-3' 3'-CGCCGG CG-5' |
When the DNA is severed, single-stranded overhangs are formed which are referred to as sticky ends due to their affinity for complementary sequences . In the experiment, the enzyme can be inactivated by heat denaturation , for example at 65 ° C. for 20 minutes.
literature
- Giedre Tamulaitiene & Virginijus Siksnys (2008): NotI is not boring. In: Structure. Vol. 16, No. 4, pp. 497-498. PMID 18400171