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目的 为了获得人的血管生成素基因，并在原核表达系统中进行表达。方法 利用ＲＴ－ＰＣＲ技术 从人的肝脏组织中扩增出Ａｎｇ基因片段，并将其克隆到ｐＧＥＭ－Ｔ载体中进行序列分析，再亚克隆到原核表达到 ｐＥＴ２８ａ（＋）载体中。结果 构建了重组表达载体ｐＥＴＡ，转化宿主菌ＢＬ２１（ＤＥ３）后，经ＩＰＩＧ诱导，Ａｎｇ基因蛋白在 ＢＬ２１（ＤＥ３）中表达了相对分子质量约为１７０００的重组蛋白。ＳＤＳ－ＰＡＧＥ和薄层扫描分析表明，外源蛋白的表达量 占菌体蛋白总量的１０．６４％。结论 Ａｎｇ基因的克隆与表达为进一步研究其生物活性及应用奠定了基础。
Objective To obtain human angiogenin gene and express it in prokaryotic expression system. Methods The Ang gene fragment was amplified from human liver tissue by RT-PCR and cloned into pGEM-T vector for sequence analysis. The fragment was subcloned into prokaryotic expression vector pET28a (+). Results The recombinant expression vector pETA was constructed and transformed into host strain BL21 (DE3). After induced by IPIG, Ang protein was expressed in BL21 (DE3) with a relative molecular weight of 17000. SDS-PAGE and TLC analysis showed that the expression of foreign protein accounted for 10.64% of the total bacterial protein. Conclusion The cloning and expression of Ang gene laid the foundation for further study of its biological activity and its application.