摘要
为了鉴定胞内劳森菌(Lawsonia intercellularis,LI)外膜蛋白LI0902在细菌感染中的作用,本试验以胞内劳森菌疫苗DNA为模板,设计特异性引物进行PCR扩增,并成功构建诱饵表达载体pGBKT7-LI0902。将通过菌落PCR和测序验证的质粒转化酵母菌株Y2H Gold后,发现LI0902能在酵母细胞中正确表达,对酵母细胞无毒性,且无自激活活性。将含有pGBKT7-LI0902的酵母菌株Y2H Gold与猪肠上皮细胞系IPEC-1 cDNA文库菌株Y187进行杂交,筛选、验证、测序后得到3个与LI0902相互作用的宿主蛋白。本试验研究了胞内劳森菌与猪肠上皮细胞相互作用,为揭示增生性肠炎的致病机理提供了线索。
To identify the role of outer membrane protein LI0902 of Lawsonia intercellularis in bacterial infection,The DNA of Lawsonia intercellularis vaccine was used as a template,specific primers were designed to be amplified by PCR and bait plasmids pGBKT7-LI0902 was constructed.Plasmids verified by colony PCR and sequencing were transformed into yeast strain Y2 H Gold.The well expression of LI0902 was confirmed by Western blot,and showed no cytotoxicity and self-activation.The yeast strains Y2 H Gold which contained bait vector pGBKT7-LI0902 was mated with the IPEC-1 cDNA library strain Y187.3 candidate interacting proteins of LI0902 were verified.These results provided good clues for interpreting Lawsonia intercellularis host-pathogen interactions and revealing the pathogenesis of proliferative enteritis.
作者
张兰桥
赖崇德
戴益民
张庆华
钟其旺
赖芬菊
ZHANG Lan-qiao;LAI Chong-de;DAI Yi-min;ZHANG Qing-hua;ZHONG Qi-wang;LAI Fen-ju(School of Biology Science and Engineering,Jiangxi Agricultural University;School of Animal Science and Technology,Jiangxi Agricultural University)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2020年第1期72-76,共5页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(31360617)
江西省自然科学基金资助项目(20161BAB204183)
江西农业大学研究生创新专项资金资助项目(NDYC2018-S004)
作者简介
张兰桥(1994-),男,硕士;通讯作者:赖芬菊,E-mail:laifenju@jxau.edu.cn。
