肝脏特异性基因疫苗研究  

Studies on Liver-specific Gene Vaccine

作　　者：张利涛[1] 刘慧琳[1] 曹以诚[1] 

机构地区：[1]华南理工大学生物科学与工程学院,广东广州510006

出　　处：《现代食品科技》2010年第6期562-565,共4页Modern Food Science and Technology

摘　　要：本研究构建了肝脏特异性表达的载体,为进一步构建肝脏特异性的siRNA载体做前期准备。通过PCR方法扩增得到人Alpha-1抗胰蛋白酶(human-α1-anti-Trypsin)启动子片段(以下简称hAAT启动子)。回收纯化后克隆到pCDNA6载体中,同时以EGFP作为标记蛋白。将构建好的载体分别转染到肝癌细胞HePG2、乳腺癌细胞MDA-MB-231和肺癌细胞A549中,验证其在肝脏中特异性表达的能力。然后在荧光显微镜下观察。结果表明:载体在肝癌细胞中能很好的表达EGFP,在肺癌细胞A549、乳腺癌细胞MDA-MB-231中不表达EGFP。说明采用human-α1-anti-Trypsin启动子可以实现目的基因在肝癌细胞中特异性表达,这项研究应用于抗肝癌、肝炎的siRNA基因治疗,将有利于显著提高治疗效果,减少对身体的副作用。Liver-special expression vector was constructed for the further preparation of the liver-special expression siRNA.The hAAT promoter was amplified by PCR,which was purified and cloned to pCDNA6 vector.EGFP was the marker protein.pCDNA6-hAAT-EGFP and pCDNA6-CMV-EGFP were transfect to HepG2,MDA-MB-231,A549 cell lines,to validate the specificity of the vector.Then,these cell lines were observed by the fluorescence microscope.Result showed that,pCDNA6-hAAT-EGFP can express EGFP in the HepG2 cell line.However,no EGFP was expressed in MDA-MB-231 and A549 cell line.The vector with hAAT promoter can achieve specific expression in HepG2 cell line.It will dramatically improved the therapeutic effect,and reduced adverse effect in siRNA treatent.

关 键 词：特异性表达 HEPG2 hAAT启动子 EGFP 

分 类 号：TQ464[化学工程—制药化工]