CLEC4M慢病毒载体的构建及其在K562细胞中的表达

• Main Author: WANG Yuanyuan
• Format: Article
• Language: zho
• Published: Editorial Department of Journal of Clinical Hepatology 2014-06-01
• Series: Linchuang Gandanbing Zazhi
• Subjects: hepertitis C virus; lentivirus infections; dendritic cells
• Online Access: http://www.lcgdbzz.org/qk_content.asp?id=5815&ClassID=42414109

ObjectiveTo construct the lentiviral vector encoding CLEC4M and prepare K-562 cells with stable overexpression of CLEC4M. MethodsThe gene sequence of normal CLEC4M was cloned by RT-PCR and then inserted into GV166 vector to construct GV166-CLEC4M lentiviral expression vector, and then lentiviral packaging was performed by transfection of 293T cells. The obtained lentiviral liquid was used to infect human leukemia cell line K-562. Real-time PCR and Western blot were used to detect the overexpression of CLEC4M in K-562 cells. ResultsSequencing showed that the recombinant lentiviral expression plasmid GV166-CLEC4M was successfully constructed. Lentiviruses could efficiently infect K-562 cells, according to real-time PCR. CLEC4M was successfully over-expressed in K-562 cells at mRNA and protein levels. ConclusionThe construction of lentiviral vector encoding CLEC4M lays a foundation for further study of CLEC4M gene involved in HCV entry into host cells.