中文摘要：

目的构建人grp78基因真核表达载体，并建立稳定高表达grp78的人宫颈癌HeLa细胞系。方法用RT—PCR方法从人宫颈癌HeLa细胞中扩增grp78基因编码区，将PCR产物克隆到pcDNA3．1（＋）真核表达载体，构建重组质粒pcDNA3．1（＋）／grp78并测序鉴定。用构建成功的pcDNA3．1（＋）／grp78真核表达载体转染入人宫颈癌HeLa细胞，经G418筛选获得grp78稳定高表达的HeLa细胞系，并用RT—PCR及Western印迹方法鉴定。结果成功构建pcDNA3．1（＋）／grp78真核表达载体，筛选获得稳定高表达人grp78的HeLa细胞系。结论grp78真核表达载体的成功构建和稳定高表达grpTS的HeLa细胞系的建立为进一步研究grp78的功能奠定了基础。

英文摘要：

Objective To construct eukaryotic expression vector of human grp78 and establish its stable transfected HeLa cell line. Methods The CDS of grp78 was amplified from HeLa cell line by RT-PCR and cloned into pcDNA3.1 （ ＋ ） . The recombinant plasmid pcDNA3.1 （ ＋ ） /grp78 was sequenced. HeLa cells were transfeeted with pcDNA3.1 （ ＋ ） /grp78 and selected with G418. The expression of grp78 was identified by RT-PCR and Western Blot. Results The eukaryot- ic expression plasmid of pcDNA3.1 （ ＋ ） /grp78 was successfully constructed and grp78 stably ex- pressing HeLa cell line was established. Conclusion The recombinant eukaryotic expression vector pcDNA3.1 （ ＋ ） /grp78 and the established Hela cell line stably expressing grp78 may be used for further study of grp78 function in vitro.