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Cloning and Expression of Genes Encoding F107-C and K88-1NT Fimbrial Proteins of Enterotoxigenic Escherichia coli from Piglets

机译:仔猪产肠毒素大肠杆菌的F107-C和K88-1NT纤维蛋白编码基因的克隆与表达

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摘要

We cloned two genes coding F107-C and K88-1NT fimbrial subunits from strains E. coli C and 1NT isolated from Thua Thien Hue province, Vietnam. The mature peptide of faeG gene from strain E. coli 1NT (called faeG-1NT) is 100 % similarity with faeG gene, while the CDS of fedA gene from strain C (called fedA-C) has a similarity of 97 % with the fedA gene. Expression of the faeG-1NT and fedA-C genes in E. coli BL21 Star™ (DE3) produced proteins of ~31 and 22 kDa, respectively. The effect of IPTG concentration on the K88-1NT and F107-C fimbriae production was investigated. The results showed that 0.5 mM IPTG is suitable for higher expression of K88-1NT subunit, while 0.75 mM IPTG strongly stimulated expression of F107-C subunit. The optimal induction time for expression was also examined. Generally, highest expression of K88-1NT subunit occurred after 6 h of induction, while that of F107-C subunit is after 14 h.
机译:我们从分离自越南Thua Thien Hue省的大肠杆菌C和1NT菌株中克隆了两个编码F107-C和K88-1NT纤维亚基的基因。大肠杆菌1NT菌株的faeG基因的成熟肽(称为faeG-1NT)与faeG基因有100%的相似性,而菌株C的fedA基因的CDS(称为fedA-C)与fedA的相似性为97%基因。 faeG-1NT和fedA-C基因在大肠杆菌BL21 Star™(DE3)中的表达分别产生约31 kDa和22 kDa的蛋白质。研究了IPTG浓度对K88-1NT和F107-C菌毛生产的影响。结果表明0.5mM IPTG适合K88-1NT亚基的更高表达,而0.75mM IPTG则强烈刺激F107-C亚基的表达。还检查了表达的最佳诱导时间。通常,K88-1NT亚基的最高表达发生在诱导6小时后,而F107-C亚基的最高表达发生在14小时后。

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