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.
出处
《食品工业科技》
CAS
CSCD
北大核心
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)
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
infectious hypodermal ＆ haematopoietic necrosis virus（IHHNV）
coat protein
Pichia Pastoris
eukaryoticexpression
作者简介
彭小莉（1980一），女，工程师，研究方向：食品工程、食品微生物、水产病毒等。
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