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目的通过原核表达获得普马拉病毒重组核蛋白。方法采用RT-PCR方法,从感染普马拉病毒的鼠肺标本中扩增得到编码核蛋白1-117 aa的基因片段。将截短的核蛋白基因片段插入原核表达载体PQE30构建重组质粒,转化至大肠杆菌M15,经IPTG诱导表达,利用N i-NTA Agarose对表达产物进行纯化。结果重组蛋白得到了高效表达。表达产物分子量15 000,在菌体中主要以可溶性蛋白形式存在。通过亲和层析,得到了纯化的重组蛋白。结论普马拉病毒重组核蛋白基因片段得到了有效表达。
Objective To obtain Purumala virus recombinant nucleoprotein by prokaryotic expression. Methods The gene fragment encoding nucleoprotein 1-117 aa was amplified by RT-PCR from rat lung samples infected with Pumala virus. The truncated nucleoprotein gene fragment was inserted into the prokaryotic expression vector PQE30 to construct a recombinant plasmid, which was transformed into E. coli M15 and induced by IPTG. The expressed product was purified by N i-NTA Agarose. Results The recombinant protein was highly expressed. The expressed product has a molecular weight of 15 000 and mainly exists as a soluble protein in the cell body. By affinity chromatography, the purified recombinant protein was obtained. Conclusion The recombinant plasmid of Pumala virus was efficiently expressed.