中文摘要：

【目的】构建ZFP580基因的原核表达质粒，转化大肠杆菌，诱导表达并分离纯化大鼠锌指蛋白ZFP580。【方法】根据ZFP580 cDNA序列的开放阅读框设计引物，利用PCR技术扩增ZFP580的开放阅读框cDNA序列，将PCR扩增的目的片段与载体pET30a分别双酶切、回收纯化后做定向连接，Nco Ⅰ、Xbo Ⅰ双酶切电泳筛选、鉴定并测序。将构建的重组质粒pET30a—ZFP580转化大肠杆菌BL21（DE3），由异丙基-β-D-半乳糖苷（IPTG）诱导表达并纯化目的蛋白。【结果】成功扩增ZFP580基因并构建了表达质粒pET30a-ZFP580，测序结果表明与GeneBank公布的序列一致。含有该质粒的大肠杆菌BL21（DE3）经IPTG诱导表达后，产物蛋白分子量约为19kDa。【结论】成功表达并纯化了大鼠ZFP580蛋白，为进一步研究锌指蛋白ZFP580的功能奠定了基础。

英文摘要：

[Objective]To construct the ZFP580 prokaryotic expression plasmid pET30a-ZFP580. Expressed ZFP580 gene in E. coli, and its protein was purified. [Methods] The primers were designed according to the open reading frame of the ZFP580 cDNA sequence, the PCR fragment and pET30a were digested with both Nco Ⅰ and Xho Ⅰ. Recombinant prokaryotic expression vector was identified by restriction enzymes digestion, agarose gel electrophoresis and DNA sequencing. The BL21 （DE3） contained the expression plasmid was induced by IPTG and the ZFP580 protein was purified. [ Results ] The open reading frame of ZFP580 gene was amplified successfully from plasmid, and its sequence was identical with that reported in GeneBank. The expression plasmid pET30a-ZFP580 was successfully constructed. The BL21（DE3） contained the expression plasmid pET30a-ZFP580 could express a 19 kDa protein after induced by IPTG and the protein was purified. [Conclusions] In this study, the protein of rat ZFP580 was expressed and purified successfully, which provided the materials for further investigating the ZFP580 protein function.