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

发布时间：2018-05-06 18:20

本文选题：重组 + 猪细小病毒　；参考：《中国兽医学报》2017年01期

【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。
[Abstract]:To explore the potential value of recombinant porcine pseudorabies virus (rPRV-VP2) strain of recombinant PPV VP2 gene as a candidate strain of attenuated virus vaccine, to study its proliferation characteristics, virus titer, genetic stability and one-step growth curve in a variety of cells, and to investigate the potential value of recombinant porcine pseudorabies virus rPRV-VP2 strain as a candidate strain of attenuated vaccine. The biological characteristics of transcription and translation were studied, and the safety of mice was preliminarily explored. The results showed that the TCID50 of the recombinant rPRV-VP2 strain on VEROSTN PK-15 and IPEC cells was 104.375 / 0.1 mL / 106.55 / 103.75 / 0.1 mL / 105.625 / 0.1 mL, respectively. The virulence of recombinant virus in St cells was about the same as that of parental PRV HB-98 strain (106.125 TCID 50% 0.1 mL), and the recombinant virus kept the integrity of PRV virus. After 20 generations of continuous transmission of the virus, the results of green fluorescence observation showed that the VP2 gene in the recombinant virus was effectively expressed and the genetic stability was good. The results of safety test showed that the recombinant virus was safe to mice. This experiment laid a foundation for the development and utilization of recombinant porcine pseudorabies virus rPRV-VP2 strain with PPV VP2 gene.
【作者单位】：河南农业大学牧医工程学院;大北农集团动物医学研究中心;
【基金】：河南省重大科技专项资助项目(111100110300)
【分类号】：S852.651

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