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Gene cloning and high expression of clostridium difficile toxin A C-terminal repeated unit

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Author:
No author available
Journal Title:
JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
Issue:
18
DOI:
10.3321/j.issn:1000-2790.2006.18.005
Key Word:
艰难梭菌;毒素A;受体结合区;克隆,分子;基因表达

Abstract: 目的:克隆并表达艰难梭菌毒素A羧基末端受体结合区 (CDTAR)基因.方法:PCR扩增CDTAR基因并将其克隆到表达载体pET-22b(+),重组质粒转化到E.coli BL21(DE3),经IPTG诱导表达,聚丙烯酰胺凝胶电泳(SDS-PAGE)对表达产物进行分析.结果:构建了含CDTAR基因的重组质粒pET-CDTAR,IPTG诱导后SDS-PAGE显示表达出Mr约为35.7 ku的重组蛋白,占菌体总蛋白的36.1%,可溶性表达占上清的22.2%,包涵体中约占24.9%.结论:成功克隆了CDTAR基因,并构建表达了CDTAR重组蛋白,为进一步研究CDTAR功能及研制艰难梭菌疫苗奠定了基础.

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