Expression of human apM1 gene lentiviral vector in 293 T cells and its significance
- VernacularTitle:人脂联素基因慢病毒载体在293 T细胞中的表达及意义
- Author:
Jie HUANG
;
Liangdi XIE
- Publication Type:Journal Article
- Keywords:
human apM1;
lentiviral vector;
293 T cells;
vascular smooth muscle cells
- From:
Chinese Pharmacological Bulletin
2003;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Aim To investigate the expression of the lentiviral vector carrying a human apM1(hapM1) gene in 293 T cells and its bioactivity.Methods The CDS region of hapM1 gene was subcloned into the lentiviral vector from a cloning vector,and the transfer plasmid(PNL-hapM1-IRES2-EGFP)of the lentiviral vector was constructed.The three plasmids of the lentiviral vector: PNL-hapM1-IRES2-EGFP,HELPER,and VSVG were cotransfected to 293 T cells to produce the recombinant lentivirus;293 T cells were infected by the recombinant lentivirus and were identified with fluorescent microscope,RT-PCR and western blot.Inhibitory effect of the recombinant hapM1 on proliferation of vascular smooth muscle cells(VSMCs) was identified.Results The sequencing results showed that the sequence of the subcloned hapM1 gene was completely identical to that reported in GeneBank.The virus titre of the recombinant hapM1 lentivirus was 2.0?105 TU?L-1.After the recombinant lentivirus infected 293 T cells,RT-PCR and Western blot results showed that the expression of hapM1 in the hapM1-293 T group could be measured.The recombinant hapM1 inhibited the proliferation of VSMCs.Conclusions The recombinant lentivirus carrying a human apM1 gene was constructed successfully.The recombinant hapM1 could be expressed in 293 T cells and had its bioactivity.
