中国野桑蚕抗病毒蛋白基因(Lipase)的克隆与活性鉴定  

Cloning of an Antiviral Protein Gene(Lipase) From Chinese Wild Silkworm,Bombyx mandarina Moore and its Bioactivity Assay

作　　者：姚慧鹏[1] 何芳青[1] 郭爱芹[1] 曹翠平[1] 鲁兴萌[1] 吴小锋[1] 

机构地区：[1]浙江大学动物科学学院,杭州310029

出　　处：《蚕业科学》2008年第3期466-471,共6页ACTA SERICOLOGICA SINICA

基　　金：国家高技术研究发展计划"863"项目(编号2007AA10Z159);国家重点基础研究发展计划"973"项目(编号2005CB121003-3);教育部新世纪人才计划项目(编号NCET-06-0524)

摘　　要：从中国野桑蚕幼虫中肠细胞克隆获得了抗家蚕核型多角体病毒BmNPV的脂肪酶基因(Lipase)cDNA(GenBank:AY945212)。该基因cDNA大小906 bp,编码301个氨基酸,蛋白质分子质量约为28.9 kD。进一步克隆了其全长基因组,结果表明该基因由2 147 bp组成,包含4个外显子和3个内含子。该基因在野桑蚕体内的表达具有组织特异性,仅限于野桑蚕中肠组织表达,且在幼虫龄中表达水平较高,而在幼虫眠期和熟蚕期几乎没有表达。通过基因体外表达获得的重组蛋白Lipase,能够有效抑制BmNPV病毒对家蚕的感染,说明该蛋白具有较强的抗BmNPV的生物学活性。A cDNA encoding an antiviral protein, Lipase （GenBank accession AY945212）, was cloned from Chinese wild silkworm, Bombyx mandarina Moore, based on the available information of silkworm, Bombyx mori. The size of its cDNA is 906 bp, encoding 301 amino acids with predicated molecular weight of 28.9 kD. Its full length of genomic sequence was also analyzed. It consists of 2 147 bp, including 4 exons and 3 introns. The gene expression analysis showed that the lipase gene was only expressed in midgut, and its expression level was higher during the ingesting stage while almost no expression during molting and mature stage. In order to identify its bioactivity, large amount of recombinant lipase was obtained by efficient gene expression system in vitro, and activity assay showed that this protein had strong antiviral activity against BmNPV.

关 键 词：野桑蚕 脂肪酶 抗家蚕核型多角体病毒蛋白 基因克隆 活性鉴定 

分 类 号：S885.9[农业科学—特种经济动物饲养] Q78[农业科学—畜牧兽医]