对虾传染性皮下及造血组织坏死病毒衣壳蛋白基因在毕赤酵母中的表达  

Expression of the coat protein gene of IHHNV in shrimp in Pichia Pastoris

作　　者：彭小莉[1,2] 谢明星[2] 刘棠[2] 陈双雅[2] 王群力[2] 陈伟玲[2] 王景明[2] 张其清[1] 

机构地区：[1]厦门大学生物医学工程研究中心,福建厦门361005 [2]厦门出入境检验检疫局,福建厦门361012

出　　处：《食品工业科技》2012年第13期67-70,共4页Science and Technology of Food Industry

基　　金：国家质检总局科技基金资助项目(2006IK015)

摘　　要：对虾传染性皮下及造血组织坏死病毒是"须向OIE申报的甲壳动物重要疾病"之一。将该病毒主要结构蛋白基因克隆至毕赤酵母穿梭表达质粒pPIC9K,构建重组表达载体(命名为pPIC9K-IV),限制性内切酶BglⅡ对其进行酶切线性化,采用电穿孔法转化到毕赤酵母GS115宿主菌。采用PCR方法分析和G418筛选来鉴定重组的毕赤酵母,诱导表达的产物分别进行ELISA分析或Western blot免疫印迹鉴定,分泌表达产物分子量大小为40ku左右,原核表达时制备的兔抗对虾传染性皮下及造血组织坏死病毒衣壳蛋白的血清可与真核表达的目的蛋白发生特异性反应。The infectious hypodermal ＆ haematopoietic necrosis virus is one of the main diseases of the penaeid shrimp. The gene of coat protein of IHHNV was constructed into Pichia Pastoris secretory expression vectors pPIC9K,named pPIC9K-IV. The recombinant plasmid pPIC9K-IV was linearized with Bgl II,and then was transformed into Pichia Pastoris GS115 by electroporation. After PCR analysis and G418 screening,the pPIC9K- IV recombinant was expressed with 0.8% （VN） methanol. The expression product was confirmed by Western blot and ELISA analysis. The results showed that the coat protein was successfully expressed,its product was about 40ku,and could recognized specifically by rabbit polyclonal anti-serum.

关 键 词：传染性皮下及造血组织坏死病毒 衣壳蛋白 毕赤酵母 真核表达 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]