Cloning and expression of placental growth factor protein of human -1 (hPLGF-1) in Rosetta E.coli expression system

Angiogenesis or new blood vessels generation is the most important factor affecting cell growth and proliferation in physiologic and pathologic conditions. Placenta Growth Factor (PLGF) is one of the important proteins for angiogenesis induction. In this study, placenta-derived PLGF-1 cDNA was cloned and expressed in Escherichia coli expression system.

In this experimental study, the human placenta-derived PLGF-1 gene was amplified using specific primers and was cloned into pET32a and pET28a expression vectors. In order to express target protein, pET32a-PLGF-1 and pET28a-PLGF-1 constructs were transformed into E.coli Rosetta. PLGF-1 expression was induced by IPTG, and then the yield of expressed protein and its solubility was analyzed by SDS-PAGE and Western blotting assay.

The pET32a-PLGF-1 and pET28a-PLGF-1 constructs were confirmed by sequencing. The bacterial lysates derived from IPTG-induced samples showed exact proteinand confirmed by western bloting. The majority of the expressed protein was insoluble.

The PLGF-1 is well expressed in Rosetta E.coli system and it could be used in different project.