貂圆环病毒Cap蛋白在毕赤酵母中的分泌表达

Secretory expression of mink circovirus capsid protein in Pichia pastoris

为研制水貂圆环病毒(MiCV)亚单位疫苗或建立相关诊断技术,利用常规PCR扩增出优化后的貂圆环病毒Cap蛋白基因序列,并插入毕赤酵母分泌型表达载体pPICZαA中。获得的重组表达载体pPICZαA-Cap以SacⅠ进行线性化后,经电击转化整合入毕赤酵母GS115菌株中,用含Zeocin的YPD平板筛选出重组子pPICZαA-Cap-GS115。PCR、SDS-PAGE和Western-blot分析鉴定的结果显示,貂圆环病毒Cap蛋白成功在毕赤酵母中分泌表达。

To develop mink circovirus（MiCV）diagnosis techniques or explore the virus subunit vaccine,the 684 bp optimized MiCV Cap gene was amplified and inserted into Pichia pastoris secretory expression vector pPICZαA.The recombinant expression vector pPICZαA-Cap was linearized by SacⅠ and transformed into P.pastoris GS115 strain by electric shock.The recombinants were screened in YPD flats containing Zeocin and identificated by PCR,SDS-PAGE and Western-blot.The results suggested that MiCVCap was successfully expressed secretly in P.pastoris.