以对虾白斑病毒DNA为模板扩增出Vp28基因，经限制性内切酶（SmaI，Not I）酶切后按正确的读码框顺序插入到pGEX-4T-2表达载体上，重组质粒转化大肠杆菌，经菌落PCR和质粒双酶切鉴定、序列测定确认，证实成功地构建了Vp28基因原核表达载体．转化菌经IPTG诱导表达，SDS-PAGE显示有与预期大小约52kD相吻合的融合蛋白带，18℃诱导24h后获得了可溶性表达的目的蛋白，采用Glutathione Sepharose 4B亲和层析对重组蛋白进行纯化，获得了纯度很高的目的蛋白．%The Vp28 gene was cloned by PCR from White Spot Syndrome Virus. After digested by Not I and Sma I, it was inserted into the plasmid pGEX-4T-2 with the right reading frame sequence to construct the expression vector, which can express the fusion protein with GST tag. The fusion protein was expressed after inducing with IPTG at 18 ℃ for 24 h and purified for antibody preparation by Glutathione Sepharose affinity chromatography.
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