Restriction generated oligonucleotides utilizing the two base recognition endonuclease CviJI*.
AUTOR(ES)
Swaminathan, N
RESUMO
The conversion of an anonymous DNA sample into numerous oligonucleotides is enzymatically feasible using an unusual restriction endonuclease, CviJI. Depending on reaction conditions, CviJI is capable of digesting DNA at a two or three base recognition sequence. CviJI normally cleaves RGCY sites between the G and C to leave blunt ends. Under 'relaxed' conditions CviJI* cleaves RGCY, and RGCR/YGCY, but not YGCR sites. In theory, CviJI* restriction of pUC19 (2686 bp) should produce 157 fragments, 75% of which are smaller than 20 bp. Instead, 96% of the CviJI* fragments were 18-56 bp long and none of the fragments were smaller than 18 bp. Thermal denaturation of these fragments generates sequence specific oligonucleotides homologous for the cognate template. The enzymatic conversion of anonymous DNA into sequence specific oligomers has implications for several conventional and novel molecular biology procedures.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=308007Documentos Relacionados
- Rapid shotgun cloning utilizing the two base recognition endonuclease CviJI.
- Molecular cloning of the three base restriction endonuclease R.CviJI from eukaryotic Chlorella virus IL-3A.
- Efficient cloning of PCR generated DNA containing terminal restriction endonuclease recognition sites.
- New restriction endonuclease CviRI cleaves DNA at TG/CA sequences.
- Specificity of substrate recognition by the EcoRI restriction endonuclease.
