对虾白斑综合症病毒结构蛋白60B的原核表达和其基本氨基酸序列的初步分析

PROKARYOTIC EXPRESSION OF VP60B OF SHRIMP WHITE SPOT SYNDROME VIRUS AND PRIMARY ANALYSIS ON ITS AMINO ACID SEQUENCE

  • 摘要: VP60B是对虾白斑综合症病毒(WSSV)中含量很少的一个结构蛋白。VP60B的一段序列与腺病毒纤维蛋白(Adenovirustype5fiberprotein)的knobdomain的一段序列具有同源性。本试验将VP60B基因克隆到原核表达系统中,在低温条件下,诱导了VP60B蛋白的表达。结果显示VP60B在该系统主要是以包含体的形式存在。原核表达的VP60B不能被鼠抗WSSV多克隆血清所识别,预示该蛋白的免疫原性较弱。通过对VP60B氨基酸序列的分析,发现有一个跨膜区,这预示着该蛋白可能位于WSSV病毒的囊膜上。

     

    Abstract: White spot syndrome virus (WSSV) from shrimp was first found in Taiwan in 1992. Since 1993, white spot syndrome disease of shrimp caused by this virus was widely spread throughout China,Asia and Pan?Pacific Ocean. It can cause 100% cumulative mortality of cultured shrimps in 3-4 days. The virus particles contain fivemajor proteins: VP28, VP26, VP24, VP19 and VP15.The mode of entry and systemic infection of WSSV in shrimp and the funotions of these virion proteins are not clear. VP60B is a kind of minor structure proteins in WSSV particles. Using interproscan to analysis on homology to VP60B, a piece of sequence of amino acid in VP60Bwas found homology with adenovirus type 5 fiber protein knob domain. In this study, thegene of VP60B had been cloned into a prokaryotic expression system, and induced to express at low temperature. The result showed that the expressed protein was existed mainly as inclusion body in this expression system. Prokaryotic expressed VP60B could not recognize by polyclone antiserum against WSSV, which suggested that this protein might be aweak immunogen. According to the analysis of the sequence of amino acid, a trans?membrane domain was found, which suggested that this protein might locate on the envelop membrane of WSSV.

     

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