表达猪细小病毒VP2蛋白的猪伪狂犬病病毒rPRV-VP2株的生物学特性  

Biological characteristics of a recombinant pseudorabies virus rPRV-VP2 expressing VP2 protein of PPV

作　　者：付朋飞[1,2] 乔涵[1] 张宇[1] 杨兴武[1] 徐朋丽 陈红英[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002 [2]大北农集团动物医学研究中心,北京100195

出　　处：《中国兽医学报》2017年第1期11-17,共7页Chinese Journal of Veterinary Science

基　　金：河南省重大科技专项资助项目(111100110300)

摘　　要：探索重组PPV VP2基因的猪伪狂犬病病毒rPRV-VP2株作为弱毒疫苗候选株的潜在价值,对其在多种细胞上的增殖特性、病毒滴度、最适培养细胞中的遗传稳定性、一步生长曲线、转录和翻译水平等生物学特性进行研究,并初步探索对小鼠安全性。结果显示:重组病毒rPRV-VP2株在VERO、ST、PK-15和IPEC细胞上的TCID50分别为104.375/0.1mL、106.5/0.1mL、103.75/0.1mL、105.625/0.1mL。重组毒与亲本毒PRV HB-98株在ST细胞中的毒力(106.125 TCID50/0.1mL)和细胞病变相当,重组病毒保持了PRV病毒的通性。连续传毒20代后的绿色荧光观察、RTPCR、Western blot试验结果均表明重组病毒中的VP2基因得到了有效表达,且遗传稳定性良好。安全性试验结果显示:重组毒对小鼠安全。本试验为后期对PPV VP2基因重组猪伪狂犬病毒rPRV-VP2株的开发利用奠定了基础。In order to investigate the potential value of a recombinant pseudorabies virus coexpress- ing VP2 protein of porcine parvovirus（PPV） as a candidate vaccine,the biological characteristics of the recombinant virus rPRV-VP2 was examined,including the growth properties and virus ti- ters on mutiple cells, the genetic stability research, the test of one-step-growth curve and gene transcription and translation levels on the optimal cell, and the preliminary safety experiment to i- dentify whether the recombinant virus rPRV-VP2 is secure to mice. The results showed that the virus rPRV -VP2 . s similar cultural and growth properties to parental pseudorabies virus HB98 strain on ST cell. The TCIDS0 of the recombination virus rPRV VP2 on VERO, ST, PK-15 and IPEC cell lines are 10^4.375/0.1mL、10^6.5/0.1mL、10^3.75/0.1mL and 10^5.625/0.1mL respectively. Genetic stability demonstrated that the recombinant virus could stably express VP2 protein of PPV after the 20th passage and visualized by green fluorescent observation,RT-PCR and Western blot. The safety test indicated that the immunization of the recommhinant virus was secure for mice. The study lays the foundation for the further development and utilization of the recombinant virus rPRV-VP2.

关 键 词：重组 猪细小病毒 VP2基因 PRV 生物学特性 

分 类 号：S852.65[农业科学—基础兽医学]