Chu Hyuk, Kang Seungha, Ha Seckho, Cho Kwangkeun, Park Sung-moo, Han Kyung-ho, Kang Sang Kee, Lee HongGu, Han Seung Hyun, Yun Cheol H, Choi Yunjaie
School of Agricultural Biotechnology, Seoul National University, Korea.
Microbiol Immunol. 2005;49(11):941-8. doi: 10.1111/j.1348-0421.2005.tb03687.x.
The most common enteric colibacillosis in neonatal and newborns is caused by enterotoxigenic Escherichia coli(ETEC). Colonization of ETEC in the small intestine is associated with adhesions using fimbriae, which is known as a specific adhesion factor and provides highly specific means for anchoring and prerequisite for an infectious agent. In the present study we have engineered Lactobacillus acidophilus to produce recombinant K99 fimbriae, which is used for the colonization to the intestine of pigs. The expression of K99 fimbrial protein was confirmed using SDS-PAGE, immunoblot and agglutination analyses. To evaluate a function of the K99 fimbrial protein, inhibition and competition tests were performed on pre-screened intestinal brush border from pigs. The tests showed that recombinant L. acidophilus, not control L. acidophilus, had a significant inhibitory effect to and competition against K99+ E. coli in a dose dependent manner. In conclusion, we demonstrated that recombinant K99 fimbriae producing L. acidophilus was able to prevent E. coli binding to intestinal brush border.
新生儿和新生仔猪中最常见的肠道大肠杆菌病是由产肠毒素大肠杆菌(ETEC)引起的。ETEC在小肠中的定植与使用菌毛的黏附有关,菌毛是一种特异性黏附因子,为病原体的锚定提供了高度特异性的方式,也是感染的先决条件。在本研究中,我们构建了嗜酸乳杆菌以产生重组K99菌毛,用于在猪肠道中的定植。使用SDS-PAGE、免疫印迹和凝集分析确认了K99菌毛蛋白的表达。为了评估K99菌毛蛋白的功能,对预先筛选的猪肠刷状缘进行了抑制和竞争试验。试验表明,重组嗜酸乳杆菌而非对照嗜酸乳杆菌对K99+大肠杆菌具有显著的剂量依赖性抑制作用和竞争作用。总之,我们证明了产生重组K99菌毛的嗜酸乳杆菌能够防止大肠杆菌与肠刷状缘结合。