Construction and identification Bifidobacterium as a delivery system of IL-10

Abstract

Objective To construct and identify a novel IL-10 delivery system by transforming a hIL-10-containing plasmid into B.longum(BL-hIL-10). Methods A plasmid vector pBADs-GFP was selected which had been built by previous test and biosynthetic hIL-10 plasmid, through double enzyme digestion and enzyme reaction,to construct and identify PBADs-hIL-10 shuttle plasmid, then to synthesis BL-hIL-10. hIL-10 was expressed and secreted into the culture supernatant of BL-hIL-10 after 0.2%L-arabinose induction in vitro as examined by Western blot, enzyme-linked immunosorbent assay(ELISA)and RT-PCR; Culture supernatants and bacterium pellets were collected after continuous culture for 12,24 and 36h,respectively.hIL-10 was expressed and secreted into the culture supernatant of BL-hIL-10 after 0.2%L-arabinose induction in vitro as examined by Western blot,enzyme-linked immunosorbent assay(ELISA)and RT -PCR;Culture supernatants and bacterium pellets were collected after continuous culture for 12,24 and 36h, respectively. Results The BL-hIL-10 bacterial strain that can stably express hIL-10 factor was successfully screened out, and the levels of hIL-10 in both supernatant and cell pellet were similarly reached maximum at 24h of culture. Conclusion BL-hIL-10 as a novel oral hIL-10 delivery system has been successfully established, which established a basis for the treatment of IBS with transgenic Bifidobacterium. Key words: Bifidobacterium longium; IL-10; Shuttle plasmid; Construction; Identification