稳定表达HLA-A33蛋白细胞系的建立  

Establishment of a Cell Line with Stable Expression of HLA-A33 Protein

作　　者：王健[1,2] 邹宁[1] 潘煦文[2] 刘思当[1] 

机构地区：[1]山东农业大学,泰安271018 [2]中国科学院微生物研究所,北京100101

出　　处：《生物技术通报》2011年第11期212-215,共4页Biotechnology Bulletin

摘　　要：旨在构建能稳定表达HLA-A33蛋白的细胞系,并观察其在细胞中的表达水平。首先克隆取得HLA-A33基因,并将其插入慢病毒载体,经酶切和测序鉴定,确定载体构建正确。通过慢病毒系统感染正常RD细胞,将HLA-A33基因整合进RD细胞的基因组。提取构建细胞系的基因组做PCR鉴定,并通过免疫荧光和蛋白免疫印迹法检测,结果显示,HLA-A33基因成功整合入RD细胞基因组中,且在重组细胞系中成功表达。该细胞系可为A33等位基因与HBV感染后的慢性化以及EV71感染的相关研究提供试验参考。The research was to construct a cell line stably expressing the HLA-A33 protein in rhabdomyosarcoma cells（RD cell）.We first obtain the HLA-A33 gene through common cloning methods and insert the gene into Lentiviral vector.The construct of the vector was further conformed by restrictional enzyme digestion analysis and DNA sequencing.This vector was transfected into common RD cells through lentivirus with Lipofectamine 2000 in order to integrate the HLA-A33 gene to RD cell genome.The genomic DNA was extracted and identified through PCR.The expression of the HLA-A33 protein was detected by Immunofluorescence assay and Western blotting assay.Results showed that the target gene was integrated into the genome of RD cells and successfully expressed.

关 键 词：RD细胞 HLA-A33 细胞系 稳定表达 

分 类 号：R392[医药卫生—免疫学]