摘要
根据GenBank上WSSV囊膜蛋白基因vp28的序列,设计并合成引物,PCR扩增得到vp28基因,成功构建重组表达载体pET22b-vp28并转化大肠杆菌BL21(DE3)。基因工程菌株37℃IPTG诱导,表达产物经Western-blot和SDS-PAGE检测显示有与预期大小32kDa相符合的目的蛋白。用Ni2+-柱纯化的目的蛋白分别直接注射螯虾和包被饲料投喂螯虾,实验结果表明vp28在大肠杆菌中的表达产物有显著提高虾体抗WSSV感染力的作用,而且注射效果更好。
A pair of primers was designed according to the sequence of vp28 gene of White spot syndrome virus （WSSV） in the GenBank. The vp28 DNA fragment was amplified by PCR and cloned into E. coli expression vector pET- 22b（＋） successfully. Then pET22b-vp28 was transformed into E. coli. After IPTG induction at 37℃, the fusion protein with 32kDa was expressed,which was conformed by Western-blot and SDS-PAGE analysis. The envelope protein VP28 purified by Ni^2＋ -column chromatography was injected into crayfish or used as a supplement to feed crayfish. The result showed that the engineered protein VP28 expressed in E.coli could improve the immunity ability of crayfish to resist WSSV infection.
出处
《中国病毒学》
CSCD
2006年第2期178-180,共3页
Virologica Sinica
