【摘要】 为了克服生物素生物合成中的抑制作用,提高生物素操纵元基因本底水平的表达,分别构建了整合表达载体pGJj01和pGJj02,通过双交叉整合的方式先后将线性化的pGJj01和pGJj02整合到枯草芽孢杆菌AS1094的染色体上,构建了枯草芽孢杆菌GJZ01,用强启动子P43-1替换掉生物素操纵元自身的启动子Pbiotin,并在bioB和bioI基因间加入强终止子和强启动子P43-2。经PCR和Southern检测表明,整合构建正确。试验可为下一步研究生物素操纵元基因表达提供良好的宿主菌素材。更多还原

【Abstract】 In order to overcome the inhibition in biotin bio-synthesize and enhance biotin operon genes expression,two integrated expression vector pGJj01 and pGJj02 were lineared,and then integrated into bacullus subtilis AS1094 to get bacullus subtilis GJZ01.Promoter of biotin operon was replaced by promoter 43-1 and the termination between bioB gene and bioI gene was reconstructed by strong promoter P43-2.GJZ01 was detected through PCR and Southern blot.This research provides a basis for further study on biotin operon gene`s expression.更多还原