表达人去唾液酸糖蛋白受体分子细胞系的建立

Establishment of Stable Cell Line Expressing Human Hepatocyte Surface Molecular ASGPR

目的构建稳定表达人肝细胞表面分子去唾液酸糖蛋白受体（asialoglycoprotein receptor，ASGPR）的细胞系。方法逆转录PCR扩增人肝组织ASGPR大亚基H1全编码序列，插入到真核表达载体plRES2EGFP中，重组质粒pIRES2EGFP／ASGPRH1转染HeLa细胞，G418筛选，RT—PCR，Western印迹及免疫荧光检测ASGPRH1的表达。结果成功构建了pIRES2EGFP／ASGPRH1重组质粒，该质粒转染HeLa细胞后，Western印迹及免疫荧光均检测到ASGPRHI蛋白的表达。结论成功建立了稳定表达人ASGPRH1的细胞系，为进一步研究ASGPR分子奠定了基础。

Objective To establish stable cell line expressing human asialoglycoprotein receptor （ASGPR） HI subunit. Methods The full length of ASGPRH1 fragment was amplified by RTPCR from human liver and inserted into eukaryotic expression vector plRES2EGFP. The obtained recombinant was transfected into Hela cells. After selective culture with G418, stably transfected cell line was established. The transcription and expression of ASGPRH1 were identified by RT-PCR, Western blotting and immunofluorescence assays. Results The eukaryotic expression recombinant pIRES2EGFP/ASGPRH1 was successfully constructed. The human ASGPRH1 protein was expressed successfully. Conclusion The stable cell line expressing human asialoglycoprotein receptor （ASGPR） H1 subunit was established.