Glycodelin基因（PAEP）重组质粒构建及法医学应用
he establishment of recombinant PAEP eukaryotic plasmid and the application in forensic medicine
目的 构建Glycodelin基因（PAEP）重组质粒，并对其在真核细胞中的表达及法医学应用进行研究。方法 提取人胎盘组织总RNA，应用 RT-PCR技术，特异性扩增PAEP基因，并将其插入到真核质粒pCMV-HA中，构建重组质粒pCMV-HA-PAEP。脂质体介导法将重组质粒转染至HEK293细胞中，免疫印迹检测蛋白表达，免疫荧光检测Glycodelin与精细胞的结合。结果 成功构建了PAEP真核表达质粒pCMV-HA-PAEP，该重组质粒可在真核细胞HEK293中高效表达，Glycodelin可对精细胞进行识别与结合。结论 构建的PAEP重组质粒在真核细胞中高表达，表达产物与精细胞的特异结合为深入研究PAEP基因在疾病中的分子机制及法医学应用奠定基础。
o establish eukaryotic expression plasmid for PAEP and to study its expression and forensic application. Method Total RNA was extracted from the placenta of human and PAEP was amplified by RT-PCR. The obtained PCR products were subcloned into plasmid pCMV-HA. HEK293 cells were transfected with purified pCMV-HA-PAEP plasmid by lipofectamine 2000, the expressed product in the cells was detected by immunoblotting technique. Overexpressed glycodelin combined with spermatozoa was analyzed by immunofluorescent method. Results A recombinant PAEP expression plasmid pCMV-HA-PAEP was successfully constructed and efficiently expressed in HEK293 cells. Spermatozoa might be recognized and bound by overexpressed glycodelin. Conclusion The recombinant pCMV-HA-PAEP was constructed and expressed successfully in eukaryotic HEK293. Overexpressed glycodelin is useful in further study on the PAEP and application in forensic medicine.
庞灏、李冯锐
基础医学生物科学研究方法、生物科学研究技术分子生物学
法医遗传学Glycodelin克隆真核表达精细胞
forensic geneticsPAEPcloningeukaryotic expressionspermatozoa
庞灏,李冯锐.Glycodelin基因（PAEP）重组质粒构建及法医学应用[EB/OL].(2013-08-23)[2025-08-16].http://www.paper.edu.cn/releasepaper/content/201308-254.点此复制
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