| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | Cloning vector pUC57 (Genscript, USA) inserted gene PI-DN9 from metagenome of the sponge QT; expression vector pET- 32a(+); strain E. coli Top10F’; strain E. coli BL21(DE3) (Invitrogen, USA); restriction enzyme EcoRI, NotI and DNA marker (Fermentas, USA); protein markers (Novagen, Netherlands; Sigma-Aldrich, USA, iNtRON Biotechnology, Korea; Affymetrix, USA); Thrombin (Novagen, The Netherlands); Skimmed milked (Difco, USA); trypsin, ɑ- chymotrypsin, thermolysin, IPTG, NBT (Sigma-Aldrich, USA). | Get A Quote |
Marine sponge is known as a “gold mine” of natural products from marine environment. Many novel bioactive compounds have been isolated from marine sponges and sponge-associated microorganisms such as antibiotics, anti-cancer compounds, protease inhibitors, etc. In this study, we selected a gene encoding protease inhibitor from metagenome of a sponge collected in Quang Tri to express in Escherichia coli (E. coli) BL21(DE3). The gene PI-DN9 encoding protease inhibitor (1.3 kb) was cut off cloning vector pUC57/PI-DN9 containing gene PI-DN9 and inserted into expression vector pET-32a(+), the recombinant vector pET-32a(+)/PI-DN9 then was transformed and expressed in the E. coli strain BL21(DE3). Results showed t... More
