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目的 构建谷胱甘肽转硫酶－血小板因子４ （ＧＳＴ－ＰＦ４）融合蛋白表达载体，并研究其编码的蛋白质在大肠杆菌中的表达。方法通过ＲＴ－ＰＣＲ方法，从ＨＬ－６０细胞中克隆ＰＦ４ ｃＤＮＡ，然后克隆至载体ｐＵＣ１９中，序列测定后把ＰＦ４基因重组入谷胱甘肽转硫酶融合基因表达载体ｐＧＥＸ－４Ｔ－３中，并用ＩＰＴＧ诱导其在大肠杆菌中表达。结果获得了ＰＦ４ ｃＤＮＡ。序列分析表明，该序列与ＧｅｎＢａｎｋ数据库中的序列一致。重组质粒酶切鉴定表明，ＰＦ４基因已正确插入到ｐＧＥＸ－４Ｔ－３中。重组融合蛋白表达载体ＧＳＴ－ＰＦ４经ＩＰＴＧ诱导表达，在ＳＤＳ－ＰＡＧＥ后得到１条蛋白表达带，相对分子质量（Ｍｒ）约为３６ ０００。结论成功地构建融合蛋白表达载体ＧＳＴ－ＰＦ４，并在大肠杆菌中获得有效表达，为进一步研究打下良好的基础。
Objective To construct a GST-PF4 fusion protein expression vector and study the expression of its encoded protein in Escherichia coli. Methods PF4 cDNA was cloned from HL-60 cells by RT-PCR and then cloned into vector pUC19. After sequencing, PF4 gene was recombined into glutathione S-transferase gene expression vector pGEX-4T-3 IPTG induced its expression in E. coli. Results PF4 cDNA was obtained. Sequence analysis showed that the sequence was consistent with the sequence in GenBank database. Restriction endonuclease digestion indicated that the PF4 gene was correctly inserted into pGEX-4T-3. The recombinant fusion protein expression vector GST-PF4 was induced by IPTG, and a protein expression band was obtained after SDS-PAGE. The relative molecular mass (Mr) was about 36,000. Conclusion The fusion protein expression vector GST-PF4 was successfully constructed and expressed efficiently in E. coli, laying a good foundation for further study.