Citation: LU Ming-Qian, DONG Rong, WEN Shun-Hua, ZHANG Wei, WANG Qiao-Zhen, HUANG Shu-Shi, CHEN Li-Mei. Detection of Recombinant Protein Expression of Formate Dehydrogenase in Single Living Escherichia Coli Cell by Laser Tweezers Raman Spectroscopy[J]. Chinese Journal of Analytical Chemistry, ;2012, 40(12): 1845-1851. doi: 10.3724/SP.J.1096.2012.20585 shu

Detection of Recombinant Protein Expression of Formate Dehydrogenase in Single Living Escherichia Coli Cell by Laser Tweezers Raman Spectroscopy

LU Ming-Qian ^1,2 ,
DONG Rong ^2,3 ,
WEN Shun-Hua ^1,2 ,
ZHANG Wei ¹ ,
WANG Qiao-Zhen ^2,3 ,
HUANG Shu-Shi ^1,2,, ,
CHEN Li-Mei ^1,,

1.
¹ Biotechnology Research Center, Kunming University of Science and Technology, Kunming 650500, China;

Corresponding author: HUANG Shu-Shi, CHEN Li-Mei,
Received Date: 5 June 2012
Available Online: 3 August 2012
Fund Project: 本文系国家自然科学基金(No.30970263)资助项目 (No.30970263)

The detection of the expression of formate dehydrogenase (FDH) recombination proteins in E.coli by molecular methods is time-consuming and hard-working and needs to destruct E.coli cells. To explore a simple and rapid method without cell destruction to detect the real-time expression of FDH recombination proteins in E.coli, Laser Tweezers Raman spectroscopy (LTRS) was used to investigate the recombinant protein expression of formate dehydrogenase (FDH) in the single living E.coli cell at different culture times following the induction with isopropyl thiogalactoside (IPTG). The result showed that the characteristic peaks corresponding to the recombinant FDH protein were gradually enhanced with an in increase in IPTG induction time, indicating the expression and the accumulation of FDH protein in recombinant E.coli cells. This result is consistent with that obtained by the analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This evidence confirms that LTRS is an effectively method for detection of the real-time expression of FDH recombination proteins in the living single E.coli cell without cell destruction.
- Escherichia coli,
- Formate dehydrogenase,
- Expression of recombinant protein,
- Single cell,
- Raman spectroscopy
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