摘要
为研究亲环蛋白A(CyPA)对日本脑炎病毒(JEV)体外增殖能力的影响,构建了真核重组质粒pcDNA3.1-CyPA,并将其转染至BHK21细胞内,接种JEV后于不同时间点收集细胞上清液,采用荧光定量RT-PCR检测各时间点的病毒含量,绘制体外增殖曲线;同时检测不同质量浓度的CyPA原核表达蛋白对JEV复制的影响。结果显示,加入CyPA原核表达蛋白组的上清中病毒含量均显著高于对照组。结果表明,CyPA可以促进JEV的体外复制,为进一步揭示CyPA在JEV感染和致病机制中的作用及JEV抗病毒药物的研发或疫苗的大量生产奠定了一定的理论基础。
To study the impact of cyclophilin A（CyPA） on the in vitro proliferative capacity of Japa- nese encephalitis virus （JEV）, an eukaryotic recombinant plasmid pcDNA3. 1-CyPA was constructed and transfected into BHK21 cells. After being inoculated with JEV,the cell supernatant was collected at diffe- rent time points,and JEV content was detected by real-time RT-PCR. The influence of prokaryotically ex- pressed CyPA protein on JEV replication was studied. In result, the virus content of supernatant in the transfection of eukaryotic vector containing CyPA gene group and the group which was added with the pro- karyotic expressed CyPA protein were significantly higher than that of the control groups. This result con- firmed that the CyPA protein has obviously effect on replication of JEV,and it provides a theoretical basis for further reveal the CyPA role in JEV infection and pathogenic mechanism, developing antiviral drugs and the development of JEV vaccines.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2015年第7期686-692,共7页
Chinese Veterinary Science
基金
四川农业大学优秀硕士学位论文培育基金项目(YS2014006)
四川省科技支撑计划项目(2012NZ0001)
关键词
日本脑炎病毒
亲环蛋白A
荧光定量
Japanese encephalitis virus
cyclophilin A
fluorescent quantitation
