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Development of Screening Method for the Soluble Recombinant Protein using β-Lactamase as a Fusion Partner  

Lee, Jae-Hun (Interdisciplinary Program for Biochemical Engineering and Biotechnology, & School of Chemical and Biological Engineering, Seoul National University)
Hwang, Bum-Yeol (Interdisciplinary Program for Biochemical Engineering and Biotechnology, & School of Chemical and Biological Engineering, Seoul National University)
Kim, Byung-Gee (Interdisciplinary Program for Biochemical Engineering and Biotechnology, & School of Chemical and Biological Engineering, Seoul National University)
Lee, Sun-Gu (Department of Chemichal Engineering, Pusan National University)
Publication Information
Korean Chemical Engineering Research / v.47, no.5, 2009 , pp. 624-629 More about this Journal
Abstract
It is the most important step to screen soluble and insoluble proteins when we attempt to improve the solubility of recombinant proteins through directed evolution approach. Here we show that the solubility of a recombinant protein in vivo can be examined by expressing the recombinant protein with beta-lactamase as a fusion partner. First we constructed an expression system which can produc a fusion protein with the C-terminal of beta-lactamase. Two soluble proteins, i.e. adenine deaminase and aspartate aminotransferase, and insoluble GlcNAc-2-epimerase were cloned into the developed expression vector, respectively. We investigated the effect of the expression of the three recombinant fusion proteins on the growth of E. coli, and confirmed that the solubilities of the recombinant proteins correlated with cell growth rates.
Keywords
Over-Expression; Inclusion Body; Antibiotic Resistance Protein; Protein Solubility; Reporter Protein;
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