人HSP 70真核载体的构建及其在胆囊癌细胞中的表达

目的构建真核表达载体pcDNA3．0-Hsp70，并在胆囊癌细胞SGC996中进行表达。方法构建真核表达载体pcDNA3．0-Hsp70，经EcoRⅠ及BarnHⅡ双酶切鉴定并测序；通过脂质体法转染SGC996胆囊癌细胞，经G418抗性筛选获得稳定表达细胞株；用流式细胞术检测其表达情况。结果酶切电泳和DNA测序证实载体构建正确；体外转染入SGC996细胞中后，可见转染细胞有Hsp70蛋白的表达。结论成功构建了pcDNA3．0-Hsp70真核表达载体，为研究Hsp70在肿瘤免治疗中的作用奠定了基础。

Construction of the Eukaryotic Expression Vector pcDNA3.0-Hsp70 and Its Expression in Human Gallbladder Cancer Cells

Objective To construct the eukaryotic expression vector pcDNA 3.0-lisp 70 and to express Hsp 70 in human gallbladder cancer cells. Methods The eukaryotic expression vector pcDNA 3.0-Hsp 70 was constructed by introducing Hsp 70 DNA fragment into the sites of EcoR I and BamH l of pcDNA 3.0-Hsp 70 vector. The plasmid was transfected into the gallbladder cancer cells using Superfect Transfection Reagent. The expression of Hsp 70 protein was detected by flow cytometry. Results The eukaryotic expression vector pcDNA 3.0-Hsp 70 was constructed and transfected successfully into gallbladder cancer cells. The Hsp 70 protein expression was observed in the transfected cells. Conclusion The recombinant expression vector pcDNA 3.0-Hsp 70 was constructed, this study laid a foundation for the further research of the function of Hsp 70 in immunotherapy for human cancer.