摘要：[Objective] This study aimed to establish a method for quantitative detection of mRNA transcriptional level of SS2 adhesive related-factors of Streptococcus suis serotype 2 （SS2） by fluorescent quantitative PCR. []Vlethod] The gene fragments en- coding SS2 adhesive related-factors MRP, FBPS and CPS2J and a housekeeping gene aroA were amplified by reverse transcription PCR from the total RNA of SS2, cloned, and sequenced. The recombinant plasmids containing the target genes were constructed, and used as templates in Real-time PCR. [Result] Dynamic curves, stan- dard curves and melting curves of the adhesive related-factors and aroA were ob- tained by the optimized Real-time PCR system. The standard curves showed a good linear relationship between template copy number and circulation number, and the correlation coefficients （FF） of the standard curves were over 0.995. Also, these as- says were highly specific a^d there was single specific melting peak for every gene. Moreover, the assays were highly sensitive and had a detection limit of 1.0×102 copies in 1 μl of initial templates. Finally, it was highly repeatable and had a coeffi- cient of variation less than 2% for intra-assay. [Conclusion] This study will provide a way to reveal the adhesion mechanism of SS2 to different host cells at molecular level.

摘要：C8orf4,也称为甲状腺癌1(thyroid carcinoma 1,TC1),最初是从甲状腺癌及其周围正常甲状腺组织克隆而来,在脊椎动物中普遍表达,具有跨物种的序列保守性。研究表明,C8orf4在肿瘤中高表达,参与各种癌细胞间信息通讯,是一种与癌症和炎症有关的新型调节因子,并通过调节NF-κB的转录增强炎症反应,但对病毒的影响知之甚少。为研究C8orf4编码蛋白对猪流行性腹泻病毒(porcine epidemic diarrhea virus, PEDV)复制的影响,将PEDV接种于Vero细胞,通过qRT-PCR和Western blot检测不同时间点PEDV对C8orf4表达的影响;设计并合成表达C8orf4基因的真核表达载体pcDNA3.1-C8orf44-His和特异性siRNA,通过过表达和抑制C8orf4表达,利用qRT-PCR和Western blot检测C8orf4对PEDV复制的影响;进一步,通过过表达C8orf4编码蛋白,明确C8orf4编码蛋白对PEDV复制周期各阶段的增殖变化。随着PEDV感染时间的增长,细胞中C8orf4表达呈上升趋势;过表达C8orf4显著抑制PEDV复制,而干扰C8orf4表达促进PEDV复制,并且C8orf4编码蛋白主要作用于PEDV复制周期的生物合成阶段。本研究表明C8orf4编码蛋白具有抑制PEDV复制的作用,为C8orf4的功能研究提供参考,为抗PEDV复制机制研究提供基础。