利用RT-PCR扩增拟南芥螯合肽合成酶(AtPCS1)基因全序列,构建原核表达载体pET28a- AtPCS1并测序.将该载体转化大肠杆菌BL21,用0.75 mmol/LIPTG诱导融合蛋白AtPCS1-His的表达,纯化该融合蛋白.用纯化的融合蛋白免疫小鼠,ELISA法测定抗血清的效价可达1:2 000.抗血清与融合蛋白及拟南芥总蛋白的western-blot结果表明:制备的抗血清具有较高的活性和特异性.The full length of AtPCS1 gene was amplified by RT-PCR.After sequencing,AtPCS1 expression plasmid pET28a-AtPCS1 was constrasted.The E.coli BL21 containing the expression plasmid was induced by 0.75 mmol/L IPTG and the fusing protein was purified.Mouse was immunized with the purified protein. The results of ELISA show that the titer of the anti-serum is about 1:2 000 and the western-blot analysis of the anti-serum with purified protein AtPCS1-His or the total protein of Arabidopsis thaliana show that the polyclonal anti-AtPCS1-His serum has high activity and specialty.甘肃省自然科学基金(3ZS042-B25-011);; 兰州理工大学博士基金(SB08200602);; 甘肃省教育厅研究生导师基金(0703-11)资
