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

GUO Xin-yu1,JIANG Feng2,ZHAO Hong-xi2,YAO Yuan-qing1 1Department of Gynecology and Obstetrics,the Chniese PLA General Hospital,Beijing 100853;2Department of Gynecology and Obstetrics,Tangdu Hospital,Fourth Military Medical University,Xi' an 710038,China  
AIM:To construct a lentiviral expression vector carrying HLA-G5.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-EF1-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.
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