中文摘要：

[目的]克隆莱茵衣藻缺铁应答基因Femu2p,构建重组表达载体p GEX-6p-1-Femu2p,优化条件使其在大肠杆菌BL21（DE3）中表达。[方法]提取莱茵衣藻CC425的总RNA后,利用RT-PCR和两次重叠延伸PCR相结合的方法获得Femu2p的全长编码区,构建原核表达载体p GEX-6p-1-Femu2p,并将其转入大肠杆菌DL21（DE3）中,利用IPTG诱导融合蛋白的表达并优化表达条件。[结果]Femu2p的全长编码区为2 904 bp,编码967个氨基酸,理论分子量为100.4 k Da,理论等电点为8.86。序列分析结果显示该蛋白属于包含多个C2H2类型锌指结构域的Ran BP2家族。IPTG诱导融合蛋白表达的最佳条件为：IPTG浓度0.25 mmol/L,诱导温度16℃,诱导时间8 h。[结论]成功克隆了Femu2p基因的全长编码区,并使其在大肠杆菌BL21（DE3）中表达。

英文摘要：

[ Objective ] In order to clone the iron - deficiency related gene Femu2p from Chlamydomonas reinhardtii, construct the recombi- nant expression vector of pGEX -6p -1 -Femu2p,and express it in E. coli BI21 （DE3）. [ Methods] The total RNA was extracted from C. reinhardtii,the target gene was obtained by using RT- PCR and overlap extension PCR,and cloned into prokaryotic expression vector pGEX- 6p- 1. The fusion protein was expressed by IPTG induction in E. coli BL21 （DE3）. [ Results ] The Femu2p CDS sequence of 2 904 bp encodes 967 amino acid residues with a predicted molecular weight of 100. 4 kDa and an isoeletric point value of 8.86. Homolo- gy sequence analysis showed that Femu2p belong to a RanBP2 -type family with several C2H2 zinc finger domains. The expression level of the fusion protein was the highest after induced by 0. 25 mmol/L IPTG for 8 h at 16℃. [ Conclusion] The CDS sequence of Femu2p gene was successfully cloned and the recombinant protein was obtained in E. coli BL21 （DE3）.