大肠杆菌FliC基因敲除后对大肠杆菌生物膜形成的影响  

Influence of FliC knock-out Escherichia coli on the biofilm formation

作　　者：喻华英[1] 马燕[1] 

机构地区：[1]塔里木大学动物科学学院,新疆阿拉尔市843300

出　　处：《中国兽医学报》2016年第8期1301-1306,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31060344)

摘　　要：利用无疤痕基因敲除方法,构建大肠杆菌FliC基因敲除突变株。基因重组PCR结果显示:在重组转化体中FliC基因完全被卡那抗性基因和CcdB替代,获得Ⅰ型突变株(△1FliC);同时利用基因突变技术,对FliC基因起始密码子进行点突变,将ATG突变为AAA,再次进行基因重组后获得Ⅱ型FliC基因沉默菌株(△2FliC)。利用96孔微量板法,分别对野毒株、△1FliC和△2FliC进行生物膜表型检测,结果显示:△1FliC和△2FliC生物膜形成能力明显低于野毒株,从而为临床上大肠杆菌病的预防及疫苗的研制提供一定的理论基础。Experimental methods of no scars knock-out and mutant were used to knock-out flagellin FliC gene of the E. coli. The recombinant PCR results showed that the recombinant transformant strains FliC gene was completely alternatived by the kanamycin resistance gene and CcdB , so got Ⅰ mutant strain （AIFliC）, then gene dot mutations method was used to mutate ATG of FliC gene start code to AAA. After one more gene recombination,type Ⅱ mutant strain FliC（△2FliC） gene in silence strain was got. Ninety six-well micro titer plate method was used respectively to de- tect wild strain,/klFliC and △2FliC biofilm phenotype. The results showed that △1FliC and A2FliC biofilm formation were significantly lower than that of the wild strain. The test provides a theoretical basis of the development of vaccines against E. coli disease in clinical.

关 键 词：大肠杆菌 FliC基因 生物膜 基因敲除 

分 类 号：S852.61[农业科学—基础兽医学]