【作者】 朱科； 陈慧； 韩玲； 杨渝珍；

【机构】 华中科技大学同济医学院生物化学与分子生物学系；

【摘要】 目的:考察野生型地衣芽抱杆菌20386表达外源基因的可能性和表达能力。为利用生物活性基因在该菌株中的表达以开发新一带基因修饰微生态制剂作出有益的探索.方法:利用大肠杆菌一枯草芽抱杆菌穿梭表达载体在大肠杆菌DHSQ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抱杆菌20386中表达,在紫外光照射下检测绿色荧光蛋白的存在.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,而在液体比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抱杆菌20386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,还需进一步研究。更多还原

【Abstract】 Objective: By detecting the fluorescence of green fluoresent protein, we could know the possibility and capability of the expression of heterologous gene in the wide-type strain. It would be benefited to study a new type of microecologic agent with gene modification.Methods: A recombinant plasmid including the gene of green fluorescent protein was constructed in E. coli DH5 α by using shuttle vector of Bacillus subtilis-Escherichia coli and was transformed into wide-type of Bacillus licheniformis by electroporation. The fluorescence of green fluoresent protein was detected by U.V.Results: Under U.V, the green fluorescence of the clone which grew in LB plate could be observed ,but the green fluorescence could not be detected in the clone which cultured in LB liquid or the cultured supernatant.Conclusion: Heterologous gene could express in wide-type (?).licheniformis but the secreted protein would be decomposed by the protease which secreted by the strain itself, we are now studying it further.更多还原