生防枯草芽孢杆菌B579芽孢形成关键基因的研究

目的]克隆生防菌芽孢形成关键基因spo0A启动子Pspo0A,并检测其特性.方法]从生防菌B579 DNA中,经PCR扩增芽孢形成关键基因spo0A启动子Pspo0A,插入载体pGFP,构建重组表达质粒pGFP-Pspo0A.结果]重组表达质粒经双酶切和PCR鉴定,证明构建正确,测序,结果与GenBank中的序列同源性为98％.结论]该方法获得了含有绿色荧光蛋白基因和芽孢形成关键基因Spo0A的重组质粒,为进一步研究生防菌B579芽孢形成规律奠定了基础.

The Research of Spore Form Key Genes of Biocontrol Bacillus subtilis B579

Objective] To clone and express the promoter of spore form key genes of Bacillus subtilis and determine the property of expressed product.Method] The promoter of spore form key genes spo0A of B.subtilis was amplified from chromosome DNA of B.subtilis by PCR and inserted into vector pGFP.The constructed recombinant secretory expression vector pGFP-Pspo0A was transformed to B579 by Electrotransformation.Result] Both restriction analysis and PCR proved that recombinant plasmid pGFP-Pspo0A was constructed correctly.Sequencing result proved that the homology of nucleotide sequence of target gene was 98% to that reported in GenBank.Conclusion] The study will lay a foundation for further study form law of biocontrol bacterium B579.