猪链球菌7型强弱毒株SBP_bac_5差异表达蛋白的鉴定  

Identification of the differential expression of bacterial extracellular solute-binding protein family 5 in Streptococcus suis 7 virulent and attenuated strains

作　　者：金家民[1,2] 王淑杰[1,2] 高明明[3] 王延群[2] 李玉明[1,2] 汪志艳[2] 石文达[4] 刘鹤[4] 姜成刚[2] 张秀英[1] 蔡雪辉[2] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/动物疫病诊断及技术服务中心,黑龙江哈尔滨150001 [3]石河子大学,新疆维吾尔自治区石河子832000 [4]动物用生物制品国家工程研究中心,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2013年第3期206-209,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省杰出青年科学基金(JC201020)

摘　　要：前期研究表明细菌胞外溶质结合蛋白家族5(SBP_bac_5)基因在猪链球菌7型(S.suis 7)型强毒株WC0711中表达量高于弱毒株M13中,推测SBP_bac_5为S.suis 7型强弱毒株差异性表达蛋白。为进一步证实该结果,本研究采用real-time RT-PCR技术在基因水平检测S.suis 7型强弱毒株中SBP_bac_5 mRNA的表达量差异,结果显示,强毒株WC0711中SBP_bac_5的mRNA表达量为在弱毒株M13中的4.59倍。将SBP_bac_5的PCR产物克隆于pET-30a中,通过E.coli BL21诱导表达,纯化后的蛋白免疫新西兰白兔制备抗血清检测S.suis强弱病毒株中SBP_bac_5的表达量差异。结果显示,重组蛋白获得高效表达,S.suis 7型强毒株WC0711中SBP_bac_5表达量是弱毒株M13的3.8倍。Our previous study showed that the extracellular solute-binding protein family 5 （SBP_bac_5） was highly expressed in Streptococcus suis 7 virulent strain WC0711 compared to the attenuated M13. To further confirm the difference, the SBP bac 5 gene was amplified from S.suis 7 genome DNA and cloned into pET-30a for expression in E.coli. In addition, the recombinant SBP_bac_5 was expressed by IPTG induction, and the polyclonal antibodies were prepared in rabbits immunized with recombinant SBP_bac_5. Furthermore, the expressions of SB_bac_5 were detected in both S.suis 7 strain WC0711 and M13 by western blot with the polyclonal antibodies, and the results showed that the expressions of SBP_bac_5 was 3.8 fold in S.suis 7 virulent strain WC0711 than that in attenuated M13, which was consistent with the results of SBP_bac_5 mRNA expression levels by 4.59 foldin virulent strain to the attenuated strain detected by real time RT-PCR.

关 键 词：猪链球菌 细菌胞外溶质结合蛋白家族5 强毒株 弱毒株 

分 类 号：S852.61[农业科学—基础兽医学]