Prohibitin 2基因参与猪圆环病毒2型PK-15细胞感染的研究  

Prohibitin 2 is involved in PCV2 infection in PK-15 cells

作　　者：蒋一凡[1,2,3] 罗旭英 吴锐 符世豪 张佳鑫 余婉婷[1,2,3] 李周勉 王乃东[1,2,3] JIANG Yi-fan;LUO Xu-ying;WU Rui;FU Shi-hao;ZHANG Jia-xin;YU Wan-ting;LI Zhou-mian;WANG Nai-dong(College of Veterinary Medicine,Hunan Agricultural University,Changsha 410128,China;Hunan Provincial Key Laboratory of Protein Engineering in Animal Vaccines,Changsha 410128,China;Research&Development Center for Animal Reverse Vaccinology of Hunan Province,Changsha 410128,China;College of Mathematics and Computer Science,Yichun University,Yichuan 336000,China)

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128 [2]兽用蛋白质工程疫苗湖南省重点实验室,湖南长沙410128 [3]兽用疫苗逆向创制湖南省工程研究中心,湖南长沙410128 [4]宜春学院数学与计算机科学学院,江西宜春336000

出　　处：《中国兽医科学》2020年第4期506-511,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31372406);湖南省教育厅重点项目(15A086);湖南省自然科学基金面上项目(2018JJ-2177);湖南省大学生研究性学习和创新性实验计划项目;国家级大学生创新创业训练计划项目(SCX1816);湖南农业大学“双一流”建设项目(SYL2019048)。

摘　　要：为探究Prohibitin2(PHB2)在PCV2感染细胞过程中发挥的作用,首先通过间接免疫荧光试验检测PCV2在PK-15细胞中的增殖变化,同时建立PHB2荧光定量PCR检测体系,然后以PCV2已知的宿主细胞互作蛋白gC1qR为对照,检测PCV2感染PK-15细胞后PHB2 mRNA的动态变化,最后鉴定PHB2 siRNA转染PK-15细胞中PCV2 Cap的表达。结果表明,PHB2在正常及PCV2感染的PK-15细胞中均有效表达。随着PCV2感染PK-15细胞,gC1qR转录水平在病毒感染3 h显著降低,感染6 h时升高,随后(12~48 hpi)转录水平下降;而PHB2转录水平在PCV2感染6 h内显著升高,随后(12~48 hpi)mRNA水平有所下降;RNA干扰PHB2对PCV2在PK-15细胞中复制起到显著的抑制作用。上述研究结果表明,PCV2可显著影响PK-15细胞中PHB2转录,而且PHB2对PCV2病毒复制具有调节作用。In order to unveil the role of Prohibitin 2(PHB2) in the process of PCV2 infection,firstly,the PCV2 proliferation in PK-15 cells was detected by indirect immunofluorescence assay.And a real-time quantitative PCR detection method for PHB2 was established,then the gC1qR(known as host cell interaction protein of PCV2) was used as a control to detect the dynamic changes of PHB2 m RNA in PCV2-infected PK-15 cells.Finally,the PCV2 Cap expression in PK-15 cells transfected with PHB2 si RNA was identified by Western-blot.The results showed that PHB2 was efficiently expressed both in normal and PCV2-infected PK-15 cells.With the PCV2 proliferation in PK-15 cells,the transcription level of gC1qR was significantly decreased at 3 h post-transfection(hpi),but increased at 6 hpi,and then decreased again(12-48 hpi).Whereas the PHB2 transcription level was significantly increased 6 hpi,and then decreased within 12-48 hpi.Moreover,PHB2 si RNA significantly inhibited PCV2 replication in PK-15 cells.Based on the present results,this study found that PCV2 could significantly affect the transcription of PHB2 in PK-15 cells,indicated that PHB2 had a regulatory effect on PCV2 replication.

关 键 词：猪圆环病毒2型 猪肾上皮细胞 gC1qR PROHIBITIN 2 荧光定量PCR 

分 类 号：S852.659.2[农业科学—基础兽医学]