Construction,package and identification of lentiviral vector for CXCR4 gene
10.11659/jjssx.01E015050
- VernacularTitle:CXCR4基因慢病毒表达载体的构建、包装及鉴定
- Author:
Lijuan XU
;
Shufang WANG
;
Yunwei ZHANG
;
Meiyan PAN
;
Yazhuo HU
;
Li YAN
- Publication Type:Journal Article
- Keywords:
lentiviral vector;
CXCR4 Gene;
293T cell
- From:
Journal of Regional Anatomy and Operative Surgery
2015;(5):473-476
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct and identify lentiviral vector pGC-FU-CXCR4 gene. Methods CXCR4 gene amplification was used by real-time polymerase chain reaction. The target gene fragments with the digested plasmids were exchange. Then the lentiviral vector pGC-FU-CXCR4 was constructed successfully. Use the constructed lentiviral vector to infect the competent escherichia coli cells. Polymerase chain reaction analysis was used to identify the cultural clones and DNA sequencing and comparative analysis were used to positive fragments. The successfully constructed plasmids had the same sequence with the target gene. Results Polymerase chain reaction tests showed that am-plified target genes were inserted in pGC-FU vectors. The electrophoresis results,digestion showed that the reconstructed plasmid was consist-ent with the theoretical fragment and the sequence result of the positive fragments were exactly the same with the target gene. Conclusion Lentiviral vectors of CXCR4 gene over-expression were successfully constructed.