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Analysis of Toxicity in Escherichia coli from the Expression of Human Purinergic Receptor $P2X_4$  

Yu, Yon-Joo (School of Biomedical Systems, Soongsil University)
Jung, Yun-A (School of Biomedical Systems, Soongsil University)
Lim, Dong-Bin (School of Biomedical Systems, Soongsil University)
Publication Information
Korean Journal of Microbiology / v.47, no.1, 2011 , pp. 7-13 More about this Journal
Abstract
In general, expression of membrane protein in Escherichia coli is very toxic to the host organism, but the mechanism for the toxicity is not clear yet. Expression of human purinergic receptor $P2X_4$ was found to be extremely toxic to the host E. coli. We examined this toxicity by isolation and analysis of less toxic mutant proteins. We could isolate 30 less toxic mutants of $P2X_4$ after hydroxylamine mutagenesis. Western blot showed that all of them produced proteins smaller than the wild type $P2X_4$. DNA sequencing of two largest mutant proteins showed that they were lost its second transmembrane domain. Localization analysis of these mutant proteins showed that they are not in cytoplasmic membrane, but in inclusion bodies. These data showed that inactive truncated $P2X_4$ is not toxic to E. coli and membrane integration and functionality of $P2X_4$ may be needed to show host toxicity.
Keywords
membrane protein; overexpression toxicity; protein expression and purification; purinergic receptor P2X;
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