白斑综合征病毒囊膜蛋白vp28基因在毕赤酵母中的表达

白斑综合征病毒(whitespotsyndromevirus,WSSV)是危害东南亚及北美洲沿岸养殖对虾的重要病原。近十年来,国内外对该病毒的形态结构、基因组全序列、宿主范围、传播途径、致病性和组织细胞特异性等作了大量的研究1-7],并已确定该病毒的侵染与其囊膜蛋白vp28有关8]。实验根据已发表的白斑综合征病毒囊膜蛋白vp28基因序列9]设计一对引物,通过PCR扩增出该囊膜蛋白的基因片段。将该片段连接到毕赤酵母表达载体pPICZ上,在大肠杆菌中筛选到含目的基因的重组质粒pPICZVP28,用电穿孔法将重组质粒转化到毕赤酵母菌株X33中,获得了转化子X33 …

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

A pair of primers was designed based on the vp 28 gene and PCR was performed to amplify the gene f rom WSSV DNA. Insert ing the DNA fragment to the yeas-t E . coli shuf fle 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 f lask at 28 e . 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 vp 28.