绿色荧光蛋白基因转化内生解淀粉芽孢杆菌的研究

TRANSFORMATION OF BACILLUS AMYLOLIQUEFACIENS WITH GREEN FLUORESCENT PROTEIN GENE

通过重叠延申法将枯草芽孢杆菌的启动子PSJ2与绿色荧光蛋白基因(gfp)的ORF连接起来,构建绿色荧光蛋白表达盒,再通过RcoR I和PstI双酶切将表达盒连接到pUS186载体上,转化解淀粉芽孢杆菌TB2菌株,得到可发出绿色荧光工程菌,工程菌对黄瓜枯萎病菌的拮抗作用与野生菌株相当。

The PSJ2 promoter cloned from Bacillus subtilis was spliced to gfp gene by overlap extension PCR. Thus the fused translational expression cassette PSJ2 - gfp was constructed. After being digested by RcoR I and Pst I, PSJ2 - gfp expression cassette was inserted into Bacillus subtilis vector pUS186 to give pUS186PGFP, which was then transformed into Bacillus amyloliquefaciens strain TB2 to give strain TB2GFP. The strain TB2GFP appeared weak green under blue light. The inhibiting effect of this strain on the growth of cucumerium, Fusarium oxyporum f. sp. , was not remarkably different from that of TB2.