鸭疫里默氏杆菌CH3株tonB1或tonB2基因缺失对生物被膜形成的影响  

Effect of tonB1 or tonB2 gene deletion on biofilm formation by Riemerella anatipestifer

作　　者：孙冰清[1,2] 苗双[2] 姜盼[2] 崔俊生[1,2] 陶敏捷 薛亚飞[2] 丁云竹 吴金节[1] 胡青海[2] 

机构地区：[1]安徽农业大学动物科技学院,安徽合肥230036 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国预防兽医学报》2016年第3期216-220,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(31072156;31272590;31472224)

摘　　要：为探究鸭疫里默氏杆菌CH3株TonB1和TonB2蛋白对其生物被膜形成的影响,本研究采用自杀质粒同源重组的方法分别构建了tonB1和tonB2基因的缺失突变株CH3△tonB1和CH3△tonB2。同时将tonB1和tonB2基因ORF克隆于大肠杆菌-鸭疫里默氏杆菌穿梭表达载体pRES中,构建回补株CH3△tonB1(pRES-TonB1)和CH3△tonB2(pRES-TonB2)。利用结晶紫染色法对野生株CH3、突变株CH3△tonB1和CH3△tonB2及其回补株的生物被膜进行测定。结果表明,与野生株CH3相比,突变株CH3△tonB1和CH3△tonB2生物被膜形成能力显著降低(p<0.05),而回补株CH3△tonB1(pRES-TonB1)和CH3△tonB2(pRES-TonB2)的生物被膜形成能力较突变株显著增强。本研究结果表明鸭疫里默氏杆菌的TonB1和TonB2蛋白与其生物被膜的形成相关。To study the effect of tonB1 or tonB2 gene deletion on the biofilm formation by Riemerella anatipestifer, the tonB1 or tonB2 gene deletion mutants of R.anatipestifer strain CH3 was constructed by homologous recombination using the recombinant suicide plasmid, respectively. TonB1 or tonB20RF was ligated into an E.coli-R.anatipestifer shuttle vector pRES, respectively, to construct complemented plasmids pRES-TonB1 and pRES-TonB2. Then the complemented plasmid was transformed into the corresponding R.anatipestifer mutant to generate the complemented strains CH3AtonB1 （pRES-TonB1） and CH3AtonB2 （pRES- TonB2）. Biofilm formation by the wild-type CH3, the mutants CH3AtonB1 and CH3htonB2, and the complemented strains was measured with crystal violet staining. The results showed that the biofilm formation by CH3AtonB1 and CH3AtonB2 was reduced significantly compared with that by the wild-type CH3, while that by the complemented strains CH3AtonB1 （pRES-TonB1） and CH3AtonB2 （pRES-TonB2） was increased significantly than that by the corresponding mutant. Therefore, our results suggested that both TonB1 and TonB2 proteins in R.anatipestifer were involved in the biofilm formation.

关 键 词：鸭疫里默氏杆菌 TonB1 TonB2 基因缺失 生物被膜 

分 类 号：S852.61[农业科学—基础兽医学]