Chaoyin Y, Wei S, Sun M, Lin L, Faju C, Zhengquan H, Ziniu Y
Biotechnology Research Center, China Three Gorges University, Yichang, Hubei, China.
J Appl Microbiol. 2007 Aug;103(2):454-61. doi: 10.1111/j.1365-2672.2006.03269.x.
The aim of this work was to investigate the effect of cry3A promoter on the expression of cry1Ac in Bacillus thuringiensis chromosome and stably enhance the production of different cry genes under the control of cry3A promoter.
The cry1Ac gene, which is specific to Lepidopteran larvae, was integrated into the chromosome of a B. thuringiensis plasmid-free and acrystalliferous strain BMB171, under the control of cry3A promoter and cry1Ac promoter, respectively. The expression of cry1Ac genes in the chromosome of host strain was investigated. The results from sodium dodecyl sulfate-polyacrymide gel electrophoresis, crystal observation and bioassay showed that either integrated with cry3A promoter (cry3Apro-cry1Ac) or with its native promoter (cry1Acpro-cry1Ac), cry1Ac gene could efficiently and stably express in the chromosome. The production of cry3Apro-cry1Ac gene was higher than that of cry1Acpro-cry1Ac gene.
The cry3A promoter enhanced the expression of cry1Ac gene efficiently either on the chromosome or on the plasmid in B. thuringiensis strain.
So far, the comparative studies on cry3A promoter and other cry promoters were carried on B. thuringiensis plasmids. This system offers an additional method for potentially improving the efficacy of B. thuringiensis insecticidal proteins efficiently, stably and safely.
本研究旨在探究cry3A启动子对苏云金芽孢杆菌染色体上cry1Ac基因表达的影响,并在cry3A启动子的控制下稳定提高不同cry基因的产量。
将对鳞翅目幼虫具有特异性的cry1Ac基因分别在cry3A启动子和cry1Ac启动子的控制下整合到苏云金芽孢杆菌无质粒且无晶体的菌株BMB171的染色体中。研究了宿主菌株染色体中cry1Ac基因的表达情况。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、晶体观察和生物测定结果表明,无论是与cry3A启动子(cry3Apro-cry1Ac)整合还是与其天然启动子(cry1Acpro-cry1Ac)整合,cry1Ac基因都能在染色体中高效稳定地表达。cry3Apro-cry1Ac基因的产量高于cry1Acpro-cry1Ac基因。
cry3A启动子在苏云金芽孢杆菌菌株的染色体或质粒上均能有效增强cry1Ac基因的表达。
到目前为止,关于cry3A启动子和其他cry启动子的比较研究是在苏云金芽孢杆菌质粒上进行的。该系统为高效、稳定且安全地提高苏云金芽孢杆菌杀虫蛋白的效力提供了一种额外的方法。