GFP为外源报告基因在地衣芽孢杆菌20386中表达的研究

Foreign Gene Expression in Bacillus Licheniformis 20386

目的 探讨外源基因在野生型地衣芽孢杆菌 2 0 386中表达的可行性。方法 利用大肠杆菌 枯草芽孢杆菌穿梭表达载体在大肠杆菌DH5α中构建含绿色荧光蛋白 (GFP)基因的重组质粒 ,通过电转化将重组质粒转入到野生型地衣芽孢杆菌 2 0 386中表达 ,通过检测绿色荧光蛋白的存在 ,考察该野生型菌株表达外源基因的可能性和表达能力。结果 在紫外光激发下 ,在固体LB平板上生长的菌落能显示绿色荧光 ,而在液体LB培养基中培养的菌体和培养上清中未能检测到绿色荧光蛋白的存在。结论 外源基因在野生型地衣芽孢杆菌 2 0 386中能够表达 ,但在液体LB中培养时 ,可能会被该菌自身分泌的蛋白酶分解 ;亦可能被稀释而无法测到荧光 。

Objective To study the possibility of expression of foreign gene in wild type of Bacillus licheniformis 20386 using green fluorescent protein (GFP) as a report gene. Methods A recombinant plasmid including the GFP gene was constructed in E. coli DH5α using shuttle vector of Bacillus subtilis Escherichia coli and was transformed into wild type of Bacillus licheniformis by electroporation. By detecting the fluorescence of GFP, we could know the possibility and capability of foreign gene expression in the wild Bacillus licheniformis 20386. Results Under irradiation of UV, the green fluorescence of the clone that grew up in LB plate could be observed, but the green fluorescence could not be detected in the clone that cultured in LB liquid medium. Conclusion Foreign gene could express in wild type Bacillus licheniformis , but the secreted protein would be decomposed by the proteinase being secreted by the strain itself.