NOVEL PLASMID MARKER RESCUE TRANSFORMATION SYSTEM FOR MOLECULAR-CLONING IN BACILLUS-SUBTILIS ENABLING DIRECT SELECTION OF RECOMBINANTS

被引:15
作者
HAIMA, P
BRON, S
VENEMA, G
机构
[1] CTR BIOL SCI,DEPT GENET,KERKLAAN 30,9751 NN HAREN,NETHERLANDS
[2] CTR BIOL SCI,DEPT MICROBIOL,9751 NN HAREN,NETHERLANDS
来源
MOLECULAR & GENERAL GENETICS | 1990年 / 223卷 / 02期
关键词
β-Galactosidase; α-complementation; Direct selection; Forced cloning; Homologous recombination; Monomer transformation;
D O I
10.1007/BF00265052
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 versatile plasmid marker rescue transformation system was developed for homology-facilitated cloning in Bacillus subtilis. It is based on the highly efficient host-vector system 6GM15-pHPS9, which allows the direct selection of recombinants by means of β-galactosidase α-complementation. The system offers several advantages over previously described cloning systems: (1) the convenient direct selection of recombinants; (2) the ability to effectively transform B. subtilis competent cells with plasmid monomers, which allows the forced cloning of DNA fragments with high efficiency; (3) the availability of 6 unique target sites, which can be used for direct clone selection, SphI, NdeI, NheI, BamHI, SmaI and EcoRI; and (4) the rapid segregational loss of the helper plasmid from the transformed cells. © 1990 Springer-Verlag.
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页码:185 / 191
页数:7
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