Prokaryotic Expression and Purification of DsRab from Dunaliella salina

Abstract

Abstract: Previous studies indicated that the DsRab transcript could be increased by salt stress. In order to study the functions of DsRab in salinity tolerance, the open reading frame(ORF)of DsRab gene was obtained through PCR. The target fragment was cloned in pGS-21a, and the recombinant plasmid pGS-21a-DsRab was transformed into E. coli BL21(DE3). The recombinant protein was induced with IPTG. Then the prokaryotic expression condition was optiminzed to harvested more supernatant recombinant protein. The products were purified by GST-Sefinose^TM Kit, and identified by SDS-PAGE and Western blot. The results showed that the recombinant expression vector pGS-21a-DsRab was constructed successfully, the molecular weight of the recombinant protein was in the expected line. Western blot analysis showed that the recombinant protein can be identified specifically by the anti-GST antibody.

Key words: Dunaliella salina, DsRab GST, Prokaryotic expression, Purification

Li Xiujuan, Chai Xiaojie, Tao Xiaoying, Zhao Huan, Cong Yuting. Prokaryotic Expression and Purification of DsRab from Dunaliella salina[J]. Biotechnology Bulletin, 2014, 0(4): 176-180.

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