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
1. [1]
  1 Jormakka M, Byme B, Lwata S. Current Opinion in Structural Biology, 2003, 13: 418-423
2. [2]
  2 Vorholt J A. Archives of Microbiology, 2002, 178(4): 239-249
3. [3]
  3 Berrios-Rivera S J, Bennett G N, San K Y. Metabolic Engineering, 2002,(4): 217-229
4. [4]
  4 XU Xian, JIA Hong-Hua, HE Bing-Fang, WEI Ping. Food and Fermentation Industries, 2007, 33: 5-8
5. [5]
  徐娴, 贾红华, 何冰芳, 韦萍. 食品与发酵工业, 2007, 33: 5-8
6. [6]
  5 Xie C A, Li Y Q. Applied Physics, 2003, 93: 2982-2986
7. [7]
  6 Huang W E, Ward A D, Whiteley A S. Environmental Microbiology Reports, 2009, 1(1): 44-49
8. [8]
  7 MENG Ling-Jing, LIN Man-Man, NIU Li-Yuan, LIU Jun-Xian, WANG He-Jian, YAO Hui-Lu. Chinese J. Anal. Chem. , 2011, 39(9): 1394-1399
9. [9]
  孟令晶, 林漫漫, 牛丽媛, 刘军贤, 王何建, 姚辉璐. 分析化学, 2011, 39(9): 1394-1399
10. [10]
  8 Chan J W, Winhold H, Corzett M H, Ulloa J M, Cosman M, Balhorn R, Huser T. Cytometry Part A, 2007, 71A: 468-474
11. [11]
  9 Xie C A, Nguyen N, Zhu Y, Li YQ. Anal. Chem. , 2007, 79(24): 9269-9275
12. [12]
  10 Huang S S, De Chen, Patrical L P , Venkata R V, Peter S, Li Y Q. Journal of Bacteriology, 2007, 189: 4681-4687
13. [13]
  11 ZHANG Jing, NIAN Hong-Juan, LI Kun-Zhi, CHEN Li-Mei. Life Science Research, 2012, 16(1): 25-32
14. [14]
  张婧, 年洪娟, 李昆志, 陈丽梅. 生命科学研究, 2012, 16(1): 25-32
15. [15]
  12 Allen C, Büttner S, Aragon A D, Thomas J A, Meirelles O, Jaetao J E, Benn D, Ruby S W, Veenhuis M, Madeo F, Margaret W W. The Journal of Cell Biology, 2006, 174(1): 89-100
16. [16]
  13 LAI Jun-Zhuo, LIU Bin, WANG Gui-Wen, TAO Zhan-Hua, HUANG Shu-Shi. Spectroscopy and Spectral Analysi, 2011, 31(2): 412-417
17. [17]
  赖钧灼, 刘斌, 王桂文, 陶站华, 黄庶识. 光谱学与光谱分析, 2011, 31(2): 412-417
18. [18]
  14 Li R, Bonham-Smith P C, King J. Canadian Journal of Botany, 2001, 79: 796-804
19. [19]
  15 Notingher I, Verrier S, Haque S, Polak J M, Hench L L. Biopolymers, 2003, 72: 230-240
20. [20]
  16 Nijssen A, SchutT C B, Heule F, Caspers P J, Hayes D P, Neumann M H, Puppels G J. Journal of Investigative Dermatology, 2002, 119: 64-69
21. [21]
  17 Krafft C, Knetschke T, Siegner A, Richard H W F, Reiner S. Vibrational Spectroscopy, 2003, 32: 75-83
22. [22]
  18 Hamada K, Fujita K, Smith N I, Kobayashi M, Inouye Y, Kawata S. Journal of Biomedical Optics, 2008, 13(4): 044027
23. [23]
  19 Pully V V, Otto C. Ramam Spectroscopy, 2009, 40(5): 473-475
24. [24]
  20 Naito Y, Toh-e A, Hiro-o H J. Raman. Spectrosc., 2005, 36(9): 837-839
25. [25]
  21 Tang H, Yao H, Wang G, Li Y Q Feng M. Optics Express, 2007, 15(20): 12708-12716
26. [26]
  22 ZHAO Chun-Li, FAN Xiu-Jun, ZHAO Bing, YU Yang, LI Qing-Zhang, HE Yan-Hong. Journal of Northeast Agricultural University, 2004, 35(4): 441-445
27. [27]
  赵春丽, 范秀军, 赵冰, 于洋, 李庆章, 郝艳红. 东北农业大学学报, 2004, 35(4): 441-445
28. [28]
  23 Chen D, Huang S S, Li Y Q. Anal. Chem., 2006, 78(19): 6936-6941
1. [1]
  Qin Tu , Anju Tao , Tongtong Ma , Jinyi Wang . Innovative Experimental Teaching of Escherichia coli Detection Based on Paper Chip. University Chemistry, 2024, 39(6): 271-277. doi: 10.3866/PKU.DXHX202309062
2. [2]
  Tianlong Zhang , Rongling Zhang , Hongsheng Tang , Yan Li , Hua Li . Online Monitoring and Mechanistic Analysis of 3,5-diamino-1,2,4-triazole (DAT) Synthesis via Raman Spectroscopy: A Recommendation for a Comprehensive Instrumental Analysis Experiment. University Chemistry, 2024, 39(6): 303-311. doi: 10.3866/PKU.DXHX202312006
3. [3]
  Zhuomin Zhang , Hanbing Huang , Liangqiu Lin , Jingsong Liu , Gongke Li . Course Construction of Instrumental Analysis Experiment: Surface-Enhanced Raman Spectroscopy for Rapid Detection of Edible Pigments. University Chemistry, 2024, 39(2): 133-139. doi: 10.3866/PKU.DXHX202308034
4. [4]
  Jingyi Chen , Fu Liu , Tiejun Zhu , Kui Cheng . Practice of Integrating Ideological and Political Education into Raman Spectroscopy Analysis Experiment Course. University Chemistry, 2024, 39(2): 140-146. doi: 10.3866/PKU.DXHX202310111
5. [5]
  Wei Peng , Baoying Wen , Huamin Li , Yiru Wang , Jianfeng Li . Exploration and Practice on Raman Scattering Spectroscopy Experimental Teaching. University Chemistry, 2024, 39(8): 230-240. doi: 10.3866/PKU.DXHX202312062
6. [6]
  Zhaoyue Lü , Zhehao Chen , Yi Ni , Duanbin Luo , Xianfeng Hong . Multi-Level Teaching Design and Practice Exploration of Raman Spectroscopy Experiment. University Chemistry, 2024, 39(11): 304-312. doi: 10.12461/PKU.DXHX202402047
7. [7]
  Liang MA , Honghua ZHANG , Weilu ZHENG , Aoqi YOU , Zhiyong OUYANG , Junjiang CAO . Construction of highly ordered ZIF-8/Au nanocomposite structure arrays and application of surface-enhanced Raman spectroscopy. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1743-1754. doi: 10.11862/CJIC.20240075
8. [8]
  Zhaoxin LI , Ruibo WEI , Min ZHANG , Zefeng WANG , Jing ZHENG , Jianbo LIU . Advancements in the construction of inorganic protocells and their cell mimic and bio-catalytical applications. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2286-2302. doi: 10.11862/CJIC.20240235
9. [9]
  Shuyu Liu , Xiaomin Sun , Bohan Song , Gaofeng Zeng , Bingbing Du , Chongshen Guo , Cong Wang , Lei Wang . Design and Fabrication of Phospholipid-Vesicle-based Artificial Cells towards Biomedical Applications. University Chemistry, 2024, 39(11): 182-188. doi: 10.12461/PKU.DXHX202404113
10. [10]
  Siyi ZHONG , Xiaowen LIN , Jiaxin LIU , Ruyi WANG , Tao LIANG , Zhengfeng DENG , Ao ZHONG , Cuiping HAN . Targeting imaging and detection of ovarian cancer cells based on fluorescent magnetic carbon dots. Chinese Journal of Inorganic Chemistry, 2024, 40(8): 1483-1490. doi: 10.11862/CJIC.20240093
11. [11]
  Shipeng WANG , Shangyu XIE , Luxian LIANG , Xuehong WANG , Jie WEI , Deqiang WANG . Piezoelectric effect of Mn, Bi co-doped sodium niobate for promoting cell proliferation and bacteriostasis. Chinese Journal of Inorganic Chemistry, 2024, 40(10): 1919-1931. doi: 10.11862/CJIC.20240094
12. [12]
  Peng GENG , Guangcan XIANG , Wen ZHANG , Haichuang LAN , Shuzhang XIAO . Hollow copper sulfide loaded protoporphyrin for photothermal-sonodynamic therapy of cancer cells. Chinese Journal of Inorganic Chemistry, 2024, 40(10): 1903-1910. doi: 10.11862/CJIC.20240155
13. [13]
  Di WU , Ruimeng SHI , Zhaoyang WANG , Yuehua SHI , Fan YANG , Leyong ZENG . Construction of pH/photothermal dual-responsive delivery nanosystem for combination therapy of drug-resistant bladder cancer cell. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1679-1688. doi: 10.11862/CJIC.20240135
14. [14]
  Xinyu ZENG , Guhua TANG , Jianming OUYANG . Inhibitory effect of Desmodium styracifolium polysaccharides with different content of carboxyl groups on the growth, aggregation and cell adhesion of calcium oxalate crystals. Chinese Journal of Inorganic Chemistry, 2024, 40(8): 1563-1576. doi: 10.11862/CJIC.20230374
15. [15]
  Ling Liu , Haibin Wang , Genrong Qiang . Curriculum Ideological and Political Design for the Comprehensive Preparation Experiment of Ethyl Benzoate Synthesized from Benzyl Alcohol. University Chemistry, 2024, 39(2): 94-98. doi: 10.3866/PKU.DXHX202304080
16. [16]
  Pei Li , Yuenan Zheng , Zhankai Liu , An-Hui Lu . Boron-Containing MFI Zeolite: Microstructure Control and Its Performance of Propane Oxidative Dehydrogenation. Acta Physico-Chimica Sinica, 2025, 41(4): 100034-. doi: 10.3866/PKU.WHXB202406012
17. [17]
  Quanliang Chen , Zhaohui Zhou . Research on the Active Site of Nitrogenase over Fifty Years. University Chemistry, 2024, 39(7): 287-293. doi: 10.3866/PKU.DXHX202310133
18. [18]
  Yongjie ZHANG , Bintong HUANG , Yueming ZHAI . Research progress of formation mechanism and characterization techniques of protein corona on the surface of nanoparticles. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2318-2334. doi: 10.11862/CJIC.20240247
19. [19]
  Chun-Lin Sun , Yaole Jiang , Yu Chen , Rongjing Guo , Yongwen Shen , Xinping Hui , Baoxin Zhang , Xiaobo Pan . Construction, Performance Testing, and Practical Applications of a Home-Made Open Fluorescence Spectrometer. University Chemistry, 2024, 39(5): 287-295. doi: 10.3866/PKU.DXHX202311096
20. [20]
  Tianlong Zhang , Jiajun Zhou , Hongsheng Tang , Xiaohui Ning , Yan Li , Hua Li . Virtual Simulation Experiment for Laser-Induced Breakdown Spectroscopy (LIBS) Analysis. University Chemistry, 2024, 39(6): 295-302. doi: 10.3866/PKU.DXHX202312049

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(275)
HTML views(21)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142