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渗透压调节基因proBA的融合表达对大肠杆菌耐高渗胁迫能力的影响 预览 被引量:6

Construction of fused osmoregulation proBA gene from a salt-tolerant mutant of Bacillus subtilis and its influence on the osmotolerance of Escherichia coli
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摘要 枯草芽孢杆菌93151的渗透压调节基因proB和proA以重叠基因的方式组织,但是表达两个单独的蛋白质ProB和ProA.通过引物设计,在抗脯氨酸反馈抑制耐盐突变菌株93151-14的proBA基因重叠区引入一个限制性酶切位点,分别扩增出proB和proA基因,并构建融合的proBA基因.SDS-PAGE分析显示有一条新的分子质量约为85kD的蛋白带出现.相对于表达未融合的proB和proA,proBA融合基因的表达明显提高了宿主菌大肠杆菌JM83胞内游离脯氨酸含量和其耐高渗胁迫能力. The osmoregulation proB and proA genes from Bacillus subtilis 93151 are overlapping genes, which encode two proteins ProB and ProA. A restriction enzyme site was inserted in the overlapping region of proB and proA genes from a salt-tolerant mutant of B. subtilis 93151, and a fusion gene was constructed by cloning proB and proA genes respectively. SDS-PAGE analysis showed that a novel protein with molecular mass of 85 kD was observed. When expressed in E.coli, enhanced intracellular concentrations of free proline and osmotolerance of the strain carrying the fusion gene were observed, compared with the control host cell harbouring a plasmid encoding the separate ProB and ProA.
作者 刘瑞杰 陈婷 曹军卫 LIU Rui-jie CHEN Ting CAO Jun-wei * (College of Life Sciences, Wuhan University, Wuhan 430072, China)
出处 《微生物学报》 CAS CSCD 北大核心 2005年第1期 23-26,共4页 Acta Microbiologica Sinica
基金 武汉大学校科研和教改项目
关键词 枯草芽孢杆菌 脯氨酸 融合基因 耐高渗胁迫能力 Bacillus subtilis, Proline, Fusion gene, Osmotolerance
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参考文献16

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