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感染对虾(斑节对虾)中白斑综合征病毒潜伏和吞噬作用激活蛋白基因的转录本分析

Transcript Analysis of White spot syndrome virus Latency and Phagocytosis Activating Protein Genes in Infected Shrimp (Penaeus monodon).

作者信息

Shekhar M S, Dillikumar M, Vinaya Kumar K, Gopikrishna G, Rajesh S, Kiruthika J, Ponniah A G

机构信息

Genetics and Biotechnology Unit, Central Institute of Brackishwater Aquaculture, 75, Santhome High Road, R.A. Puram, Chennai, Tamil Nadu India.

出版信息

Indian J Virol. 2012 Dec;23(3):333-43. doi: 10.1007/s13337-012-0119-7. Epub 2012 Nov 27.

Abstract

Viral latency has been recently observed to be associated with White spot syndrome virus (WSSV) infection in shrimp. In the present study, shrimp samples (Penaeus monodon) surviving WSSV infection were examined for presence of WSSV in latent phase. Virus latency was observed in shrimp which were either experimentally challenged with WSSV and survived the infection or those which survived the natural infection. Three viral transcripts (ORFs 427, 151, 366) associated with latency were analyzed by real-time PCR. The shrimp surviving the natural WSSV infection on estimation with RT-PCR were found to have low grade of WSSV infection (less than 56 copies of WSSV). All the shrimp samples were RT-PCR negative for structural protein genes of WSSV, VP24 and VP28, indicating that these samples were harboring latent phase virus. RT-PCR of all the shrimp samples which survived WSSV infection revealed amplification of phagocytosis activating protein (PAP) gene (435 bp) with higher gene expression levels in experimentally challenged shrimp when compared to naturally infected shrimp. The expression of PAP in WSSV infected shrimp samples indicates its possible role in host response for resistance against WSSV infection. PAP was cloned and expressed as recombinant protein for protection studies. Shrimp were injected with three doses (5, 15 and 20 μg g(-1) body weight) of recombinant PAP. Relative percent survival of 10 % was observed in shrimp immunized with the dose of 15 μg g(-1) body weight of recombinant PAP. The expression of both WSSV latency associated and PAP genes obtained from shrimp surviving the WSSV infection, indicates the possible role of these genes in host-pathogen interaction.

摘要

最近观察到病毒潜伏与对虾的白斑综合征病毒(WSSV)感染有关。在本研究中,对感染WSSV后存活的对虾样本(斑节对虾)进行了检测,以确定其在潜伏期是否存在WSSV。在经WSSV实验性攻毒并存活下来的对虾以及自然感染后存活的对虾中均观察到了病毒潜伏现象。通过实时PCR分析了与潜伏相关的三种病毒转录本(开放阅读框427、151、366)。经RT-PCR估计,自然感染WSSV后存活的对虾WSSV感染程度较低(WSSV拷贝数少于56个)。所有对虾样本的WSSV结构蛋白基因VP24和VP28的RT-PCR检测均为阴性,表明这些样本携带的是潜伏阶段的病毒。对所有感染WSSV后存活的对虾样本进行RT-PCR检测,结果显示吞噬作用激活蛋白(PAP)基因(435 bp)有扩增,与自然感染的对虾相比,实验性攻毒的对虾中该基因的表达水平更高。PAP在WSSV感染的对虾样本中的表达表明其在宿主抵抗WSSV感染的反应中可能发挥作用。为进行保护研究,将PAP克隆并表达为重组蛋白。给对虾注射三种剂量(5、15和20 μg g(-1)体重)的重组PAP。用15 μg g(-1)体重剂量的重组PAP免疫的对虾观察到相对存活率为10%。从感染WSSV后存活的对虾中获得的WSSV潜伏相关基因和PAP基因的表达,表明这些基因在宿主-病原体相互作用中可能发挥作用。

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