Construction of a new food-grade expression system for Bacillus subtilis based on theta replication plasmids and auxotrophic complementation
Construction of a new food-grade expression system for Bacillus subtilis based on theta replication plasmids and auxotrophic complementation
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基于theta复制质粒和营养缺陷互补构建新型枯草芽孢杆菌食品级表达系统
DOI：
10.1007/s00253-007-1035-4
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发表时间：
2007-06
影响因子：
5
通讯作者：
Chen, Wei
中科院分区：
文献类型：
--
作者：
Zhao, Jianxin;Zhang, Hao;Tian, Fengwei;Ding, Xiaolin;Xia, Yu;Chen, Wei
基于可复制的食品级表达质粒和营养缺陷型互补，构建了一种新的枯草芽孢杆菌食品级表达系统。食品级B。通过从B的染色体上敲除dal基因座产生枯草杆菌宿主FG 01。subtilis 168.将一种新的θ型芽孢杆菌复制子与B重组，构建了两个食品级表达质粒pXFGT 03和pXFGT 05。pXFGT 05是从pXFGT 03中删除两个开放阅读框（ORF）而得到的。在用pXFGT 03或pXFGT 05转化FG 01后，在Luria-Bertani琼脂平板上通过质粒编码的dal基因补充宿主表型，所述dal基因用作重组体的食品级选择标记。结果表明，两个ORF的缺失对质粒的复制没有影响。将报告基因bgaB分别克隆到pXFGT 03和pXFGT 05中，置于P43启动子的控制下，并在该食品级表达系统中成功表达。对两个重组质粒的分离稳定性进行了考察，结果表明它们是完全稳定的。
A new food-grade expression system was constructed for Bacillus subtilis based on replicative food-grade expression plasmids and auxotrophic complementation. The food-grade B. subtilis host FG01 was created by knockout of the dal locus from the chromosome of B. subtilis 168. Two food-grade expression plasmids pXFGT03 and pXFGT05 were constructed by combining a novel theta-type Bacillus replicon with the B. subtilis endogenous gene dal and P43 promoter; while pXFGT05 was derived from pXFGT03 by deletion of two open reading frames (ORFs) from the original replicon. Upon transformation of FG01 with pXFGT03 or pXFGT05, the host phenotype was complemented on Luria-Bertani agar plates by the plasmid-coded dal gene, which served as a food-grade selection marker for recombinants. Results showed that deletion of the two ORFs had no impact on plasmid replication. A reporter gene bgaB was cloned into pXFGT03 and pXFGT05, respectively, under control of the P43 promoter, and it was successfully expressed in this food-grade expression system. Segregational stabilities of two recombinant plasmids were investigated, and they were fully stable.
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影响因子：
2.6
作者：
Nguyen, HD;Nguyen, QA;Schumann, W
通讯作者：
Schumann, W
DOI：
10.1016/s0021-9258(17)39775-2
发表时间：
1984-07
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
P. Z. Wang;R. Doi
通讯作者：
P. Z. Wang;R. Doi
DOI：
10.1007/bf00330534
发表时间：
2004
期刊：
Molecular and General Genetics MGG
影响因子：
--
作者：
R. Muller;T. Ano;T. Imanaka;S. Aiba
通讯作者：
R. Muller;T. Ano;T. Imanaka;S. Aiba
DOI：
10.1016/s1096-4959(02)00183-5
发表时间：
2003-01-01
影响因子：
2.2
作者：
Peng, Y;Huang, Q;Zhang, YZ
通讯作者：
Zhang, YZ
DOI：
10.1016/s0021-9258(18)93614-8
发表时间：
1968-03
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
R. Berberich;M. Kaback;E. Freese
通讯作者：
R. Berberich;M. Kaback;E. Freese