猪源ETEC的黏附素基因克隆与原核表达Gene Cloning and Prokaryotic Expression of Adhesin in Enterotoxigenic Escherichia coil from Swine
徐刚;高继业;郎静宇;唐妤;李阁;李继祥;
摘要(Abstract):
[目的]克服传统方法制备黏附素抗原的缺点。[方法]应用PCR对猪源性产肠毒素性大肠杆菌(ETEC)的黏附素F4、F5和F6的质粒中扩增出faeG、fanC和fasG基因片段,克隆测序并与pET 32a(+)构建重组表达载体。将重组表达载体转化E.coli表达菌株BL21(DE3),并分析其免疫原性。[结果]重组表达载体经IPTG诱导获得高效表达。血凝抑制试验表明,鼠抗血清能抑制标准的ETEC强毒株凝集红细胞,抑制效价在1∶128以上。这说明克隆表达的猪源ETEC黏附素蛋白具有良好的免疫原性。[结论]该研究可为进一步开发抗体制剂奠定基础。
关键词(KeyWords): 产肠毒素性大肠杆菌;黏附素;克隆;原核表达;免疫原性
基金项目(Foundation):
作者(Author): 徐刚;高继业;郎静宇;唐妤;李阁;李继祥;
Email:
DOI: 10.13989/j.cnki.0517-6611.2012.24.009
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