白斑综合征病毒囊膜蛋白vp28基因在毕赤酵母中的表达  

Expression of the envelope protein gene vp28 of the white spot syndrome virus in Pichia pastoris

作　　者：李方[1] 叶巧真[1] 何建国[1] 王莉真[1] 

机构地区：[1]中山大学生命科学学院生物防治国家重点实验室,广东广州510275

出　　处：《水产学报》2003年第5期491-494,共4页Journal of Fisheries of China

基　　金：国家自然科学基金项目(30000128);科技部"973"环境胁迫对养殖生物抗病力的影响及人工调控项目(G1999012010);国家高技术研究发展计划(863计划)(2001AA621030)

摘　　要：A pair of primers was designed based on the vp28 gene and PCR was performed to amplify the gene from WSSV DNA. Inserting the DNA fragment to the yeast-E.coli shuffle vector pPICZ and the recombinant plasmid (pPICZVP28) that contains the target DNA fragment was obtained in the E.coli strain.The pPICZVP28 was introduced into Pichia pastoris strain X33 by electroporation. The transformant strain X33-1 was grown in BMGY media in fled flask at 28℃. Induced by methanol for 72h, the samples of cell pellets and supernatant were collected by centrifugation and analyzed by SDS-PAGE and Western-blot, which confirmed that the strain X33-1 can express the WSSV’s envelope protein vp28.A pair of primers was designed based on the vp28 gene and PCR was performed to amplify the gene from WSSV DNA. Inserting the DNA fragment to the yeast-E.coli shuffle vector pPICZ and the recombinant plasmid (pPICZVP28) that contains the target DNA fragment was obtained in the E.coli strain.The pPICZVP28 was introduced into Pichia pastoris strain X33 by electroporation. The transformant strain X33-1 was grown in BMGY media in fled flask at 28℃. Induced by methanol for 72h, the samples of cell pellets and supernatant were collected by centrifugation and analyzed by SDS-PAGE and Western-blot, which confirmed that the strain X33-1 can express the WSSV's envelope protein vp28.

关 键 词：白斑综合征病毒 囊膜蛋白 斑节对虾 毕赤酵母 表达 

分 类 号：S945.47[农业科学—水产养殖] Q786[农业科学—水产科学]