不同培养基对粪肠球菌基因表达及黏附细胞能力的影响  

Influence of culture media on the expression levels of Enterococcus faecalis genes and the adhesion to IPEC-JZ cells

作　　者：冯艳红[1] 赵娣[1] 张留君 刘玲玲[1] 陈丽颖[1] 王亚宾[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002

出　　处：《江苏农业学报》2016年第4期854-859,共6页Jiangsu Journal of Agricultural Sciences

基　　金：河南省重大攻关项目(112101110100);郑州市重点实验室基金项目(114PYZX509)

摘　　要：为探讨粪肠球菌(E.faecalis)N10菌株在血清肉汤培养基和TSB培养基中不同生长时期15种毒力基因mRNA的表达情况及对IPEC-J2细胞黏附能力的影响,采用分光光度计测量N10菌株在血清肉汤培养基和TSB培养基中的OD值,绘制生长曲线,确定生长时期(对数中期、对数后期和稳定期),用RT-PCR方法检测N10菌株中不同基因不同生长时期的mRNA转录水平,然后将不同培养基不同时期的细菌黏附IPEC-J2细胞,平板计数细菌黏附数量。结果显示,血清肉汤培养基培养的细菌浓度低于TSB培养基培养的细菌浓度(P<0.01)。在血清肉汤培养基中,仅有fsrC、sprE、gelE和ant基因的最高表达量低于在TSB培养基中的,其他基因的最高表达量均高于在TSB培养基中的。黏附试验中对数中期细菌的黏附数量最多,血清肉汤培养基培养的细菌黏附数量比TSB培养基培养的细菌黏附数量少(P>0.05),但比不加血清的肉汤培养基培养的细菌黏附数量多(P<0.01)。可见,血清肉汤培养基有利于菌毛相关基因的表达,肉汤培养基中加入血清可促进细菌黏附。The mRNA expression levels of 15 virulence genes of Enterococcus faecalis strain N10 cultured in the broth with serum and TSB medium and the adhesion to IPEC-J2 cells were detected by RT-PCR and spectrophotometer. The bacteria concentration in the broth with serum was lower than that in the TSB culture medium( P < 0. 01). In broth medium with serum,the maximum expression levels of fsr C,spr E,gel E and ant genes were lower than those in TSB culture medium. The amount of bacteria adhesive to IPEC-J2 cells was the greatest at mid-log phase,and the adhesive bacteria in broth medium with serum was less than those in TSB( P > 0. 05),but more than that in serum-free broth medium( P < 0. 01).It was indicated that broth culture medium with serum was better for gene expression,and the serum conducive to bacterial adhesion.

关 键 词：粪肠球菌 毒力基因 RT-PCR IPEC-J2细胞 

分 类 号：S852.611[农业科学—基础兽医学]