doi:

DOI: 10.3724/SP.J.1096.2013.30066

Chinese Journal of Analytical Chemistry (分析化学) 2013/41:12 PP.1789-1794

Detection of Expression of Broad Bean 14-3-3b Soluble and Inclusionbody Protein in Single Living Escherichia coli Cell by Laser Tweezers Raman Spectroscopy


Abstract:
To explore a simple and rapid method without cell destruction to detect the real-time expression levels of the soluble and inclusionbody proteins in Escherichia. coli, Raman Laser Tweezers spectroscopy (LTRS) was used to investigate the structural differences and expression levels of the soluble and inclusionbody proteins of broad bean (Vicia faba L.) 14-3-3b in E. coli cells induced under different temperatures. The results showed that 14-3-3b soluble and inclusionbody protein had different Raman lines, indicating that LTRS could be used to identify the soluble and inclusionbody protein of 14-3-3b. Raman lines in 1002, 1451 and 1665 cm-1 of the soluble protein was enhanced evidently in the recombinant E. coli induced at 16℃ and were reduced significantly in the recombinant E. coli induced at 28℃. In contrast, Raman lines in 900 and 1446 cm-1 of the inclusionbody protein were decreased evidently in the recombinant E. coli induced at 16℃ and were increased significantly in the recombinant E. coli induced at 28℃. Changes in the soluble and inclusionbody protein of 14-3-3b expression levels reflected by these Raman lines was consistent with those obtained by the analysis of sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). This evidence confirms that LTRS is an effectively and rapid method for detection of the expression levels of soluble and inclusionbody proteins in the living single E. coli cell.

Key words:Escherichia coli,14-3-3b isoform,Soluble protein,Inclusion body protein,Laser tweezers Raman spectroscopy

ReleaseDate:2015-04-19 11:07:45



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