摘要
为了探究寡肽结合蛋白OppA在副猪格拉菌(Glaesserella parasuis,GPS)突破猪呼吸道上皮屏障过程中的作用,采用自然转化法构建副猪格拉菌HPS5-SQ的oppA缺失株ΔoppA,并通过测定ΔoppA在不同培养基中的生长曲线、黏附猪气管上皮细胞(swine tracheal epithelial cells,STEC)、穿透猪呼吸道上皮屏障、损伤紧密连接蛋白等能力来判定OppA对GPS定殖和突破猪呼吸道上皮屏障的影响。结果显示:通过PCR和Western blot验证,试验成功构建了ΔoppA,表达的重组OppA具有结合血红素功能;ΔoppA和HPS5-SQ野生株在TSB培养基中生长速度相当,黏附STEC能力没有差异;在加入血红素DMEM中,野生株的生长速度比ΔoppA快,对猪呼吸道上皮屏障破坏和紧密连接蛋白损伤能力比ΔoppA强。结果表明:OppA摄取血红素促进GPS生长,进而加速猪呼吸道上皮屏障损伤。
To investigate the role of OppA in the process of breaking through porcine respiratory epithelial barrier for Glaesserella parasuis(G.parasuis),the oppA deletion strainΔoppA was constructed by natural transformation.The growth curve in different media,and the abilities of adhesion,crossing porcine respiratory epithelial barrier and damaging tight junction proteins of theΔoppA were determined to evaluate the influence of oppA in breaking through the porcine respiratory epithelial barrier.The results showed that the oppA deletion strainΔoppA was successfully constructed which was verified by PCR and Western blot.The recombinant OppA had the function of binding hemes.Compared with the wild-type strain HPS5-SQ,the growth rate ofΔoppA in the TSB medium and the adhesion ability on STEC were equal;but the wild-type strain grew faster than theΔoppA in DMEM with hemes.The abilities of the wild-type strain taking hemes and breaking through porcine respiratory epithelial barrier were stronger thanΔoppA.In conclusion,OppA took in hemes,promoted the growth of GPS,and broke through the porcine respiratory epithelial barrier.
作者
刘明星
张鹏云
陈敏
蔺辉星
周红
范红结
LIU Mingxing;ZHANG Pengyun;CHEN Min;LIN Huixing;ZHOU Hong;FAN Hongjie(College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)
出处
《畜牧与兽医》
北大核心
2022年第6期97-103,共7页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(31872480)
南京农业大学大学生创新训练项目(202117XX07,202117XX22)
江苏高校优势学科建设工程专项资金项目(PAPD)。
