DOI: 10.3724/SP.J.1096.2013.30066
Chinese Journal of Analytical Chemistry (分析化学) 2013/41:12 PP.1789-1794
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.
ReleaseDate：2015-04-19 11:07:45
王中江, 江连洲. 食品工业科技, 2012, 11(33): 63-66
1 WANG Zhong-Jiang, JIANG Lian-Zhou. Science and Technology of Food Industy, 2012, 11(33): 63-66
黄庶识, 赖钧灼, 梁裕芬, 韦俊彬. 分析化学, 2011, 39(4): 521-527
2 HUANG Shu-Shi, LAI Jun-Zhuo, LIANG Yu-Fen, WEI Jun-Bin. Chinese J. Anal. Chem., 2011, 39(4): 521-527
余多慰, 袁 生. 分析测试学报, 2004, 23, (5): 63-66
3 YU Duo-Wei, YUAN Sheng. Journal of Instrumental Analysis, 2004, 23, (5): 63-66
4 Chan J W, Winhold H, Corzett M H, Ulloa J M, Cosman M, Balhorn R, Huser T. Cytometry Part A, 2007, 71A: 468-474
卢明倩, 董 蓉, 温顺华, 张 韦, 王巧贞, 黄庶识, 陈丽梅. 分析化学, 2012, 40(12): 1845-1851
刘 凡, 李美英, 徐碧玉. 广西农业科学, 2008, 39 (3) : 273-274
5 Xie C A, Nguyen N, Zhu Y, Li Y Q. Anal. Chem., 2007, 79(24): 9269-9275
6 LU Ming-Qian, DONG Rong, WEN Shun-Hua, ZHANG Wei, WANG Qiao-Zhen, HUANG Shu-Shi, CHEN Li-Mei. Chinese J. Anal. Chem., 2012, 40(12): 1845-1851
严馨蕊, 鲍永利, 董学斌, 柳忠辉. 细胞与分子免疫学, 2001, 17(4): 365-367
于在江, 马学恩, 周建华. 生物技术, 2007, 17(3): 46-48
7 LIU Fan, LI Mei-Ying, XU Bi-Yu. Guangxi Agricultural Sciences, 2008, 39 (3) : 273-274
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
8 YAN Qin-Rui, BAO Yong-Li, DONG Xue-Bin, LIU Zhong-Hui. J. Cell Mol. Immunol., 2001, 17(4): 365-367
周殿凤, 柯惟中, 衡 航, 籍 康. 光散射学报, 2006, 18(3): 241-247
9 YU Zai-Jiang, MA Xue-En, ZHOU Jian-Hua. Biotechnology, 2007, 17(3): 46-48
刘文涵, 杨 未, 张 丹. 光谱学与光谱分析, 2008, 28(2): 344-345
10 LAI Jun-Zhuo, LIU Bin, WANG Gui-Wen, TAO Zhan-Hua, HUANG Shu-Shi. Spectroscopy and Spectral Analysi, 2011, 31(2): 412-417
11 ZHOU Dian-Feng, KE Wei-Zhong, HANG Zi, JI Kang. The Journal of Light Scattering, 2006, 18(3): 241-247
12 Notingher I, Verrier S, Haque S, Polak J M, Hench L L. Biopolymers, 2003, 72: 230-240
13 Nijssen A, Schut T C B, Heule F, Caspers P J, Hayes D P, Neumann M H, Puppels G J. Journal of Investigative Dermatology, 2002, 119, 64-69
14 Krafft C, Knetschke T, Siegner A, Richard H W F, Reiner S. Vibrational Spectroscopy, 2003, 32: 75-83
15 LIU Wen-Han, YANG Wei, ZHANG Dan. Spectroscopy and Spectral Analysi, 2008, 28(2): 344-345