【摘要】 构建了原核表达质粒pET 30a(+) sBLyS ,转化大肠杆菌BL2 1(λDE3) ,IPTG诱导表达的重组蛋白以可溶部分和包涵体两种形式存在 ,为了提高可溶部分重组蛋白的比例 ,确定 16℃为诱导表达的温度 ,最佳表达时间为 12h。在此条件下大规模诱导表达 ,经Ni2 +亲和层析获得重组人sBLyS。重组蛋白的等电点约为 7 1～ 7 3,活性状态时是非共价连接的同源三聚体。MTT法测重组人sBLyS对B淋巴细胞的增殖作用结果表明 ,B细胞经抗IgM原血清活化后 ,它的增殖程度在一定的重组人sBLyS浓度范围内随其浓度的增加而提高 ,sBLyS刺激时间对B细胞增长也有显著影响 ,sBLyS以 2 μg mL刺激 3dB淋巴细胞的增殖达最大更多还原

【Abstract】 The prokaryotic expression plasmid pET-30a(+)/sBLyS was constructed and transformed into E.coli BL21 (λDE3).The recombinant protein was found to be highly expressed by the plight of soluble part and inclusion body.For the sake of enhancing the proportion of the soluble part,inducement at 16℃ for 12h was ascertained.The expressing product was then purified by Ni 2+ affinity chromatography gel.PI of the recombinant human sBLyS(rhsBLyS)is about 7.1～7.3 and it assembles into a homotrimer.The effect of rhsBLyS on B lymphocytes by MTT method told us the B lymphocytes’ proliferating capacity dose depended on concentration and also stimulating time of the rhsBLys.With rhsBLyS(2μg/mL)stimulating 3 days,B lymphocytes can proliferate the most.更多还原