采用PCR方法扩增对虾白斑病毒核糖核苷酸还原酶基因,插入到pGEX-4T-2表达载体上构建出带有目的基因的重组质粒pGEX-4T-2RR,然后将重组质粒转化大肠杆菌.转化菌经IPTG诱导后大量表达重组蛋白,通过降低诱导温度获得可溶性表达的重组蛋白,采用Glutathione Sepharose 4B亲和层析对重组蛋白进行纯化,获得了高纯度的目的蛋白.%In this study,ribonucleotide reductase gene of WSSV was amplified by PCR,digested by Sma I and Not I,and inserted into the expression vector pGEX-4T-2 to construct recombinant plasmid.Colonies with recombinant plasmid were picked for PCR and culture identified by enzyme digestion.The results show that ribonucleotide reductase gene is successfully inserted into the pGEX-4T-2 expression vector.After induced with IPTG at 18 ℃,the fusion protein is purified with Glutathione Sepharose 4B.
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