Guo Xin-yu, Jiang Feng, Zhao Hong-xi, Yao Yuan-qing
Department of Gynecology and Obstetrics, the Chniese PLA General Hospital, Beijing 100853, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2012 Jan;28(1):81-3.
To construct a lentiviral expression vector carrying HLA-G5.
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.
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.
This research completed the package of lentivirus vector encoding HLA-G5 as a tool for further study.
构建携带HLA - G5的慢病毒表达载体。
通过限制性内切酶Nhe I/Not I酶切和T4DNA连接酶连接，将HLA - G5基因的编码区克隆到慢病毒载体中。转化至感受态大肠杆菌细胞后，通过PCR鉴定候选克隆，并利用脂质体2000转染肾细胞系293T细胞以产生慢病毒颗粒，随后测定病毒滴度。
成功构建了用于HLA - G5的慢病毒载体pCDH - CMV - MCS - EF1 - copGFP载体，上清液中的病毒滴度达到了TU/mL。
本研究完成了编码HLA - G5的慢病毒载体包装，作为进一步研究的工具。
