SMALL MOBILIZABLE MULTIPURPOSE CLONING VECTORS DERIVED FROM THE ESCHERICHIA-COLI PLASMIDS PK18 AND PK19 - SELECTION OF DEFINED DELETIONS IN THE CHROMOSOME OF CORYNEBACTERIUM-GLUTAMICUM

被引：2297

作者：

SCHAFER, A ^{[1
]}

TAUCH, A ^{[1
]}

JAGER, W ^{[1
]}

KALINOWSKI, J ^{[1
]}

THIERBACH, G ^{[1
]}

PUHLER, A ^{[1
]}

机构：

[1] DEGUSSA AG ASTA MED AG,D-33790 HALLE,GERMANY

来源：

GENE | 1994年 / 145卷 / 01期

关键词：

BROAD-HOST-RANGE CONJUGATION; KANAMYCIN RESISTANCE; SEQUENCING VECTOR; LACZ; GENE DISRUPTION AND REPLACEMENT; SACB GENE; CORYNEFORM BACTERIA;

D O I：

10.1016/0378-1119(94)90324-7

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

Here we describe small mobilizable vectors based on the Escherichia coli plasmids pK18 and pK19. We combined the useful properties of the pK plasmids (e.g., multiple cloning site, lacZ alpha fragment, sequencing with M13 primers) with the broad-host-range transfer machinery of plasmid RP4 and a modified sacB gene from Bacillus subtilis. The new pK derivatives can be transferred by RP4-mediated conjugation into a wide range of Gram(-) and Gram(+) bacteria, and should facilitate gene disruption and allelic exchange by homologous recombination. As an application example, the generation of a defined deletion of the hom-thrB genes in the chromosome of the Gram(+) bacterium Corynebacterium glutamicum glutamicum is presented.

引用

页码：69 / 73

页数：5

共 22 条

[1] CONSTRUCTION AND CHARACTERIZATION OF NEW CLONING VEHICLES .2. MULTIPURPOSE CLONING SYSTEM [J].