[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).

CONCLUSION

The Bifidobacterium Infantis-mediated sKDR gene transferring system was constructed successfully and it could remarkably inhibit the proliferation of vascular endothelial cells.