Cloning and Expression of crtE from Erwinia uredovora in Escherichia coli

Abstract

Abstract: Gene crtE of Erwinia uredovora, encoding the GGPP synthase, was amplified by PCR, and cloned into pET-15b expression vector. The recombinant plasmid was transformed into E.coli to construct engineering bacterium. Overexpression of recombinant GGPP synthase was achieved in engineering bacterium by IPTG induction. The expression level was up to 42% of the total cellular proteins. The recombinant protein was found mainly in inclusion bodies, after being solved in 8 mol/L urea and refolded. The recombinant GGPP synthase in inclusion bodies was purified on a Ni2+ chelating resin column. The purified protein with a N-terminal His-tag shows a molecular weight of 34 kDa and pI of 6.3.

Key words: Erwinia uredovora, crtE, GGPP synthase, expression, purification

WANG Jing-Chao, SUN Dong-Ping, DU Gui-Cai, GUO Dao-Sen, LI Rong-Gui. Cloning and Expression of crtE from Erwinia uredovora in Escherichia coli[J]. FOOD SCIENCE, 2007, 28(7): 276-279.

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Cloning and Expression of crtE from Erwinia uredovora in Escherichia coli

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