THE ENDO-BLUE METHOD FOR DIRECT CLONING OF RESTRICTION-ENDONUCLEASE GENES IN ESCHERICHIA-COLI

被引：32

作者：

FOMENKOV, A ^{[1
]}

XIAO, JP ^{[1
]}

DILA, D ^{[1
]}

RALEIGH, E ^{[1
]}

XU, SY ^{[1
]}

机构：

[1] NEW ENGLAND BIOLABS INC,BEVERLY,MA 01915

来源：

NUCLEIC ACIDS RESEARCH | 1994年 / 22卷 / 12期

关键词：

D O I：

10.1093/nar/22.12.2399

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A new E.coli strain has been constructed that contains the dinD1::LacZ(+) fusion and is deficient in methylation-dependent restriction systems (McrA(-), McrBC(-), Mrr(-)). This strain has been used to clone restriction endonuclease genes directly into E.coii. When E.coli cells are not fully protected by the cognate methylase, the restriction enzyme damages the DNA in vivo and induces the SOS response. The SOS-induced cells form blue colonies on indicator plates containing X-gal. Using this method the genes coding for the thermostable restriction enzymes Taql (5'TCGA3') and Tth111l (5'GACNNNGTC3') have been successfully cloned in E.coli. The new strain wilt be useful to clone other genes involved in DNA metabolism.

引用

页码：2399 / 2403

页数：5

共 33 条

[1]

BIRNBOIM HC, 1979, NUCLEIC ACIDS RES, V7, P1513

[2]

CROSA JH, 1977, GENE, V2, P95

[3]

Heitman J, 1993, Genet Eng (N Y), V15, P57

[4] SOS INDUCTION AS AN INVIVO ASSAY OF ENZYME-DNA INTERACTIONS [J].