不同荧光蛋白基因标记利迪链霉菌

 利迪链霉菌（Streptomyces lydicus）A01 是一株分离自北京郊区对植物病原真菌具有广谱抑制作用的放线菌，在植物真菌病害防治中具有良好的应用前景。为了检测红霉素启动子在利迪链霉菌A01 中的活性，为后期对菌株A01 进行遗传改造提供技术支持，同时对生防菌A01 进行遗传标记以研究和阐明其在生态环境中的生物学行为规律，本实验采用两亲本接合的方法，将增强绿色荧光蛋白（EGFP）和红色荧光蛋白（RFP）基因片段克隆到携带红霉素启动子（ermE^*）的链霉菌表达载体pIB139 中，成功构建了以egfp 和rfp 为报告基因的重组载体pIB139-EGFP 和pIB139-RFP，并转化利迪链霉菌A01，突变株在荧光显微镜下观察到较强的绿色荧光和红色荧光，同时PCR 鉴定结果正确。这表明重组载体pIB139-EGFP 和pIB139-RFP 成功转入菌株A01，并且ermE^*启动子成功启动egfp 和rfp 基因表达。

Different Fluorescent Protein Gene to Label Streptomyces lydicus A01

Streptomyces lydicus A01 is isolated from the soil of the suburban vegetable field in Beijing (China), and it shows a strong inhibitory activity against several plant pathogenic fungi. In order to detect the activity of the erythromycin promoter (ermE^*) in Streptomyces lydicus A01, and to obtain the labeled strain for further research, the egfp 720 bp and rfp 678 bp segments are inserted into the expression vector pIB139 separately, the recombinant vectors pIB139-EGFP and pIB139-RFP are successfully obtained. The pIB139-EGEP and pIB139-REP are transformed into Streptomyces lydicus A01 strain by intergeneric conjugation. Transformants are identified by the fluorescence observation with microscope. It is shown that the two recombinant vectors are successfully integrated into the strain A01, and that the ermE^* promoter could induce egfp and rfp gene expression in Streptomyces lydicus A01.