摘要
目的构建癌胚抗原(CEA)与热休克蛋白70(HSP70)的真核双表达质粒,并检测其在体外的表达。方法从结肠癌及正常肝组织中经RTPCR扩增获得CEA及HSP70cDNA,并将其定向克隆至真核双表达质粒pIRES中,重组质粒pIRESCEA及pIRESCEAHSP70,经酶切及测序鉴定后,分别转染仓鼠卵巢细胞(CHO);经RTPCR及ELISA法检测,CEA和HSP70在CHO细胞中得到有效表达。结果真核双表达载体pIRESCEAHSP70构建成功且CEA和HSP70在CHO细胞中得到有效表达。结论真核表达载体pIRESCEAHSP70的成功构建为治疗CEA阳性肿瘤提供一定的实验基础。
Objective To construct the eukaryotic eoexpression plasmid encoding carcinoembryonic antigen（CEA） and heat shock proteins 70 （HSP70）, then detect the expression of the plasmid in vitro. Methods cDNA encoding CEA and HSP70 were obtained by RT-PCR amplifications from colon cancer and normal liver respectively and cloned into an eukaryotic expression plasmid internal ribosome entrysite（plRES）. The constructed plasmids plRES-CEA and plRES-CEA-HSP70 were confirmed by restriction enzymolysis and DNA sequencing. Chinese hamster ovary cells（CHO） were transformed by plasmid pIRES-CEA and pIRES- CEA-HSP70 respectively. RT-PCR and ELISA identify the cDNA of CEA and HSP70 could be co-expressed efficiently in CHO cells. Results The eukaryotic expression plasmid pIRES-CEA-HSP70 was constructed successfully and the plasmid could co-express CEA-HSP70 efficiently in CHO cells. Conclusion The successful construction of eukaryotic expression plasmid pIRES-CEA-HSP70 establishs the experimental base for the treatment of cancer with CEA positive.
出处
《福建医科大学学报》
2005年第3期260-264,共5页
Journal of Fujian Medical University
基金
福建省自然科学基金资助项目(C0410043)
关键词
癌胚抗原
热休克蛋白质类
疫苗
DNA
carcinoembryonic antigen
heat-shock proteins
vaccines, DNA
