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https://dspace.ctu.edu.vn/jspui/handle/123456789/71457
Title: | Expression of a novel gene encoding protease inhibitor from metagenome of sponge in Vietnam |
Authors: | Tran, Thi Hong Ton, That Huu Dat Nguyen, Phuong Hoa Pham, Viet Cuong Nguyen, Thi Kim Cuc |
Keywords: | Escherichia coli Expression vector Protease inhibitor Recombinant protein Sponge |
Issue Date: | 2018 |
Series/Report no.: | Academia journal of biology;Vol. 40, No. 04 .- P.77-84 |
Abstract: | 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-DN9encoding protease inhibitor (1.3 kb) was cutoff cloning vector pUC57/PI-DN9 containing gene PI-DN9and 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 that recombinant protein (50kDa) was expressed successfully at 25°C, 1 mM of IPTG in 5 hours. The recombinant protein was purified using Ni-NTA affinity chromatography column. Western blot assay and bioactive assay showed good activity of the purified protein. |
URI: | https://dspace.ctu.edu.vn/jspui/handle/123456789/71457 |
ISSN: | 2615-9023 |
Appears in Collections: | Academia journal of biology |
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