对虾白斑综合征病毒囊膜蛋白VP110在中国明对虾鳃细胞中结合蛋白的鉴定与特性
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中国海洋大学 海水养殖教育部重点实验室, 山东 青岛 266003

作者简介:

赵建梅(1977-), 女, 博士研究生, 研究方向为水生生物免疫学. E-mail:zjm69318@126.com

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S945

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国家973计划项目(2012CB114405); 国家科技支撑计划(2012BAD17B01); 国家公益性行业(农业)科研专项(201103034).


Interaction between WSSV envelope protein VP110 and binding proteins on the gills of Chinese shrimp, Fenneropenaeus chinensis
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Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao 266003, China

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    摘要:

    在中国明对虾, pET-32(a)+1VP110, 蛋白。以作为诱饵蛋白实验结合蛋白质谱分析鉴定结合蛋白结果显示中国明对虾鳃细胞中的肌动蛋白和精氨酸激酶PCRAK, pGEX-4T-1通过实验进一步证实发生结合。克氏原螯虾, rAK中国明对虾在人工感染荧光定量检测结果显示基因表达水平显著上调时达到峰值然后下降至正常水平酶底物法检测结果同样显示鳃细胞中酶活性在感染后发生显著上调。本研究旨在为深入了解囊膜蛋白在感染宿主过程中的作用提供基础依据。

    Abstract:

    White spot syndrome virus (WSSV) is one of the most virulent viral pathogens of the shrimp-farming industry worldwide, causing high mortality and resulting in serious economic losses. The viral envelope proteins play very important roles in WSSV infection of shrimps. To investigate the interaction between WSSV envelope protein VP110 and gill cell proteins of the Chinese shrimp, , a truncated VP110 protein containing the RGD motif was recombinantly expressed in BL21(DE3) as a fusion protein with Trx/His/S-tag. Using a pull-down assay, two prominent protein bands with molecular weights of 40 kD and 42 kD were identified from gill cell proteins of using recombinant VP110 (rVP110), which were identified as arginine kinase (AK) and was cloned and expressed as a fusion protein with GST-tag using the pGEX-4T-1 vector, and the binding interaction between the recombinant AK (rAK) and rVP110 was further confirmed by a pull-down assay. In an neutralization assay, rAK appeared to be able to partially block WSSV infection and delayed the death of WSSV-infected crayfish. In addition, following WSSV infection, mRNA level in the gills was upregulated compared with the control group, as assessed by real-time quantitative PCR. The AK enzyme activity in the gills was also upregulated. These results suggested that AK in plays a role in WSSV infection.

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赵建梅,唐小千,战文斌.对虾白斑综合征病毒囊膜蛋白VP110在中国明对虾鳃细胞中结合蛋白的鉴定与特性[J].中国水产科学,2014,21(1):10-18
ZHAO Jianmei, TANG Xiaoqian, ZHAN Wenbin. Interaction between WSSV envelope protein VP110 and binding proteins on the gills of Chinese shrimp, Fenneropenaeus chinensis[J]. Journal of Fishery Sciences of China,2014,21(1):10-18

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  • 在线发布日期: 2015-06-30
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