[Cloning and prokaryotic expression of Bifidobacterium infantis-mediated sKDR and its effect on proliferation of vascular endothelial cells].
OBJECTIVE
To construct Bifidobacterium Infantis-mediated sKDR gene transferring system and to investigate its effect on the proliferation of vascular endothelial cells.


METHODS
sKDR gene amplified through PCR, and pET32a plasmid extracted from E. coli JM109 were digested respectively by two kinds of restriction enzyme (EcoR I and Xho I) and then were connected by T4 DNA Ligase. Finally, the recombinant plasmid was transformed into Bifidobacterium Infantis by electroporation. Human umbilicus vein endothelial cells (HUVECs) were cultivated in the nutritive media containing the extract of positive transformed bacteria for 24 h. The cell viability was analyzed with MTT assay.


RESULTS
The positive transformed Bifidobacterium Infantis with recombinant pET32a-sKDR plasmid was established and could express sKDR at the levels of gene and protein. Compared with the untreated group, the proliferation of HUVECs cultivated with the extract of positive transformed bacteria was inhibited significantly (P<0.01).


CONCLUSIONS
The Bifidobacterium Infantis-mediated sKDR gene transferring system was constructed successfully and it could remarkably inhibit the proliferation of vascular endothelial cells.
Mao
Shu-Hua
SH
Department of Abdominal Cancer, West China Hospital, Sichuan University, Chengdu, China.
Ji
Ling-Ling
LL
Liu
Hong
H
Xu
Yi-Zhou
YZ
Huang
Huang
H
Huang
Ying
Y
Yi
Cheng
C
D001644:Bifidobacterium / Q000235:genetics* / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018274:Electroporation; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000096:biosynthesis* / Q000235:genetics
D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D040301:Vascular Endothelial Growth Factor Receptor-2