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小反刍兽疫病毒P基因的克隆及其结构与功能分析 被引量:7

Cloning of the Phosphoprotein Gene of Peste des Petits Ruminants and Analysis of Its Structure and Function
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摘要 【目的】对小反刍兽疫病毒P基因进行克隆,并对其结构和功能的关系进行分析,为研究小反刍兽疫病毒P蛋白的功能及其在病毒增殖过程中的作用奠定基础。【方法】利用RT-PCR的方法对小反刍兽疫P基因全长进行克隆,通过生物学软件对其核苷酸序列和氨基酸序列进行了比对,利用I-TASSER进行三级结构预测,然后分析其结构和功能之间的关系。【结果】P基因和MV、CDV、RPV、DMV和PDV的P基因的相似性分别为62.9%、63.3%、64.4%、65.1%和60.4%,氨基酸的相似性分别为45.9%、46.2%、50.6%、50.3%和47.0%。三级结构表明该蛋白由3个结构域构成,中间是由多个α-螺旋构成的结构域,主要行使与L大蛋白和RNA结合的功能,两边分别是N-端和C-端结构域,C-端结构域含有3个α-螺旋,分别位于458—468aa、492—499aa和503—505aa位,主要作为N蛋白α-螺旋的结合位点。【结论】本研究成功地克隆小反刍兽疫病毒P基因,分析和预测了p蛋白的结构和功能的关系,为其进一步的生物学功能研究及其应用奠定了基础。 【Objective】 The objective of the present study was to clone the gene of PPRV phosphoprotein and analyze the structure-function relationship of phosphoprotein.【Method】 The phosphoprotein entire gene of PPRV was cloned by RT-PCR,in addition,the nucleotide and a mino acid sequences were analyzed and compared with other members of Morbillivirus through bioinformatics methods,and the protein tertiary structure was predicted using I-TASSER.【Result】 Sequence analysis showed that the homologies of nucleotide sequences between PPRV and MV,CDV,RPV,DMV and PDV were 62.9%,63.3%,64.4%,65.1%,and 60.4%,respectively,and the homologies of the deduced amino acid sequences were 45.9%,46.2%,50.6%,50.3%,and 47.0%,respectively.The tertiary structure and analysis indicated that the phosphoprotein was composed of three distinct structural domains:an N-terminal hydrophilic domain,a central domain,and a C-terminal hydrophilic domain.In addition,the central hydrophobic domain of phosphoprotein had lots of alpha helix with the potential to bind both of the large protein and RNA.The C-terminal hydrophilic domain was composed of three alpha helix,and they lay in 458-468aa,492-499aa and 503-505aa,respectively,and might play a role in their binding of alpha helix of the nucleoprotein.【Conclusion】 The phosphoprotein gene from PPRV was successfully cloned.Furthermore,the structure and function of this gene were analyzed and predicted.
出处 《中国农业科学》 CAS CSCD 北大核心 2010年第11期2355-2362,共8页 Scientia Agricultura Sinica
基金 农业公益性行业科研专项(200803026) 甘肃省国际合作项目(0805WCGA133)
关键词 小反刍兽疫病毒 P蛋白 克隆 结构与功能 分析 PPRV phosphoprotein cloning structure and function analysis
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