Simple approach for oligonucleotide‐directed mutagenesis of any double‐stranded circular DNA

FEBS Letters

Drutsa, V.L.¹  Kaberdin, V.R.¹ 

[1] A.N. Belozersky Laboratory of Molecular Biology and Bioorganic Chemistry, Moscow State University, Leninskiye Gory, Moscow 119899, USSR

关键词: Oligonucleotide-directed mutagenesis;    Pribnow box;    Nick-translation;   

DOI  :  10.1016/0014-5793(91)80594-S

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A new site-directed method for introducing mutations into any region of plasmid vector close to the unique restriction site is described. It is based on the use of 5′-phosphorylated mutagenic and nonphosphorylated auxiliary oligonucleotides and a specific combination of enzymatic procedures including ‘nick-translation’ as a key step. The method efficiency was demonstrated by constructing the deletion-insertion mutation wich creates the consensus Pribnow box in a promoter-testing plasmid. The yield of the target mutation was up to 85–95%.

【 授权许可】

Unknown