A GENERAL-METHOD FOR RAPID SITE-DIRECTED MUTAGENESIS USING THE POLYMERASE CHAIN-REACTION

被引：674

作者：

LANDT, O ^{[1
]}

GRUNERT, HP ^{[1
]}

HAHN, U ^{[1
]}

机构：

[1] FREE UNIV BERLIN,INST KRISTALLOG,ABT SAENGER,TAKUSTR 6,W-1000 BERLIN 33,GERMANY

来源：

GENE | 1990年 / 96卷 / 01期

关键词：

mutagenesis; PCR; protein and generic engineering; ribonuclease T1; Taq polymerase;

D O I：

10.1016/0378-1119(90)90351-Q

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

We have developed a general and rapid method for site-directed mutagenesis usign primed amplification by the polymerase chain reaction. Starting rom a doubl-stranded DNA template, this method requires only one simple specific mutagenic primer and two universal sequencing primers flanking the region to be mutated further upstream and downstream, respectively. Two test the method, two different mutants of the RNase T1-encoding gene have been constructed by this technique. Twelve sequenced mutant clones all showed the expected mutations without any wild-type background. © 1990.

引用

页码：125 / 128

页数：4

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