人类α型叶酸受体基因的克隆及其真核表达载体的构建

目的 探讨如何以人类α型叶酸受体(FOLR-1)基因为基础构建核酸疫苗。方法 应用RT-PCR技术，从人类卵巢癌细胞系-SKOV3细胞中扩增FOLR-1基因，插入克隆载体pGEM-T Easy，经DNA自动测序仪测序证实后，以亚克隆法构建于真核表达载体pcDNA3．1( )，并使用限制性内切酶酶切鉴定。结果 从卵巢癌细胞系SKOV3中成功扩增出FOLR-1基因，并克隆人pcDNA3．1( )载体。结论 成功构建FOLR-1的重组克隆及真核表达载体，为今后利用FOLR-1进行卵巢上皮性肿瘤的免疫及导向治疗研究打下了基础。

Cloning of gene of human folate receptor alpha and its construction of eukaryotic expression vector

Objective To study how to construct polynucleotide vaccine based 013.the gene of human folate receptor alpha(FOLR-1).Methods FOLR4 gene was amplified frcrn human ovarian carcinoma cell SKOV3 by RT-PCR.The product of PCR was ligated into pGEM-T Easy vector after restriction endonuclease digestion.Gene fragment encoding FOLR-1 was correctly ins~ted into the vector,which Was determined by an automated DNA sequencer and restriction endonuclease digestion,and then Was subdoned tO corresponding siteS cut with EcoR I plus Xho I of eukaryotic expression vector pcDNA3.1(+).Results FOLR-1 gene(774 bp)Was successfully oanstructed.And the gene sequence cloned into pcDNA3.1(+)was consistent with the known sequence after determination.Conclusion The recombinant human FOLR-1 gene clone has been established successfully and thus provides a basis for further research of the immunotherapy or targeting therapy for ovarian carcinoma.