人血管生成素基因的克隆与表达

Cloning and Expression of Human Angiogenin Gene

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 the gene of human angiogenin and express it in procaryotic expression system.Methods A fragment of human angiogenin (Ang) gene was amplified from liver tissue by RT- PCR, cloned into pGEM T vector and suquenced, then subcloned into procaryotic expression vector PET28a( + ) The recombinant plasmid pFTA was transformed to E . coli B121 (DE3) and expressed under the induction of IPTG. Results The recombinant protein with a molecular weight of about 17000 was expressed. SOS - PAGE and thin layer scanning proved that the expressed product contained 10.64%of the total somatic protein. Conclusion The study laid a foundation of further study on the biological activity and application of human angiogenin.