Construction of novel shuttle expression vectors for gene expression in Bacillus subtilis and Bacillus pumilus.
Construction of novel shuttle expression vectors for gene expression in Bacillus subtilis and Bacillus pumilus.
复制标题
DOI：
10.2323/jgam.61.124
复制
发表时间：
2015-08
期刊：
影响因子：
--
通讯作者：
H. Shao;Q. Cao;Hong-Yan Zhao;X. Tan;H. Feng
中科院分区：
文献类型：
--
作者：
H. Shao;Q. Cao;Hong-Yan Zhao;X. Tan;H. Feng
通过枯草芽孢杆菌菌株S1-4的基因组测序检测天然质粒（pSU 01）。构建了两个基于pSU 01的穿梭表达载体pSU 02-AP和pSU 03-AP，使得能够在B中稳定复制。枯草杆菌WB 600。这些载体含有报告基因aprE，其编码来自短小芽孢杆菌BA 06的碱性蛋白酶。表达载体pSU 03-AP仅含有最少的复制元件（rep、SSO、DSO），结构稳定性较好，表明pSU 01中的其余基因（ORF 1、ORF 2、mob、hsp）对B中质粒的结构稳定性不是必需的。枯草芽孢杆菌此外，用拷贝数估计为每条染色体100个以上的pSU 03-AP更有效地实现了碱性蛋白酶的重组生产。此外，pSU 03-AP还可用于在B中的转化和复制。pumilus BA 06进行选择性加压培养。因此，pSU 03-AP有望成为枯草芽孢杆菌和B中基因表达的有效工具。pumilus。
A native plasmid (pSU01) was detected by genome sequencing of Bacillus subtilis strain S1-4. Two pSU01-based shuttle expression vectors pSU02-AP and pSU03-AP were constructed enabling stable replication in B. subtilis WB600. These vectors contained the reporter gene aprE, encoding an alkaline protease from Bacillus pumilus BA06. The expression vector pSU03-AP only possessed the minimal replication elements (rep, SSO, DSO) and exhibited more stability on structure, suggesting that the rest of the genes in pSU01 (ORF1, ORF2, mob, hsp) were unessential for the structural stability of plasmid in B. subtilis. In addition, recombinant production of the alkaline protease was achieved more efficiently with pSU03-AP whose copy number was estimated to be more than 100 per chromosome. Furthermore, pSU03-AP could also be used to transform and replicate in B. pumilus BA06 under selective pressure. In conclusion, pSU03-AP is expected to be a useful tool for gene expression in Bacillus subtilis and B. pumilus.