人类白细胞抗原-G5慢病毒表达载体构建与病毒包装  

Construction of eukaryotic expression vector and package of lentivirus vector encoding human leukocyte antigen-G5

作　　者：郭新宇[1] 姜锋[2] 赵宏喜[2] 姚元庆[1] 

机构地区：[1]解放军总医院妇产科,北京100853 [2]第四军医大学唐都医院妇产科,陕西西安710038

出　　处：《细胞与分子免疫学杂志》2012年第1期81-83,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30973200)

摘　　要：目的:构建人类白细胞抗原G5(HLA-G5)的表达载体,并进行慢病毒包装,为进一步研究HLA-G5功能提供基础工具。方法:人工合成HLA-G5全长序列经测序正确后,定向接入真核表达载体pCDH-CMV-MCS-EF1-copGFP,转化入大肠杆菌,酶切鉴定并测序正确后进行慢病毒包装和滴度测定。结果:经琼脂糖凝胶电泳鉴定、PCR鉴定及测序,HLA-G5表达质粒质量合格,序列比对与设计序列符合率100%,HLA-G5慢病毒滴度测定为7.06×108。结论:成功地构建了HLA-G5表达载体并完成了慢病毒包装,为今后的研究工作提供了基础工具。AIM： To construct a lentiviral expression vector carrying HLA-GS. METHODS： The CDs region of HLA-G5 gene was cloned into the lentiviral vector by restriction endonuclease Nhe I/Not I digestion and T4DNA ligase ligation. After transformation into completent E. coli cells, the candidate clones were identified by PCR and kidney cell line 293T cells by lipofectamine 2000 to produce the lentiviral particles, and the viral titer was determined. RESULTS： The lentiviral vector pCDH-CMV-MCS-EFI-copGFP Vector for HLA-G5 was constructed successfully, and the virus in the supernatant reached a titer of TU/mL. CONCLUSION： This research completed the package of lentivirus vector encoding HLA-G5 as a tool for further study.

关 键 词：人类白细胞抗原G5 真核表达载体 慢病毒 

分 类 号：R392[医药卫生—免疫学]