单细胞激光拉曼光谱检测重组大肠杆菌细胞表达甲酸脱氢酶

引用本文: 卢明倩, 董蓉, 温顺华, 张韦, 王巧贞, 黄庶识, 陈丽梅. 单细胞激光拉曼光谱检测重组大肠杆菌细胞表达甲酸脱氢酶[J]. 分析化学, 2012, 40(12): 1845-1851. doi: 10.3724/SP.J.1096.2012.20585 shu

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

单细胞激光拉曼光谱检测重组大肠杆菌细胞表达甲酸脱氢酶

卢明倩 ^1,2, ,
董蓉 ^2,3, ,
温顺华 ^1,2, ,
张韦 ^1, ,
王巧贞 ^2,3, ,
黄庶识 ^{1,2,
,} ,
陈丽梅 ^{1,
,}

通讯作者: 黄庶识,陈丽梅; 黄庶识,陈丽梅

基金项目:
本文系国家自然科学基金(No.30970263)资助项目 (No.30970263)

摘要: 甲酸脱氢酶(FDH,EC1.2.1.2)在工业生产中有重要的应用价值,工业上应用的FDH可以通过构建高水平表达重组FDH蛋白的基因工程菌生产,用分子生物学的方法检测重组蛋白的高效表达和积累操作繁杂,耗时耗力且需要破碎细胞。为了寻找一种简单快速,不需破碎细胞,且能实时检测FDH重组蛋白在基因工程菌中表达情况的方法,本研究应用单细胞激光拉曼光谱分析技术(LTRS),研究IPTG诱导不同时间后甲酸脱氢酶重组蛋白(FDH)在大肠杆菌细胞中的表达水平。结果表明,FDH的特征峰1004,1355,1455和1667 cm^-1随着IPTG诱导时间的延长而增强,说明在诱导培养过程中FDH重组蛋白在重组大肠杆菌细胞中表达并积累,这4个峰强的增加值所反映的FDH表达量与SDS-PAGE电泳分析结果一致。实验结果证明,LTRS是快速有效检测单个大肠杆菌活细胞体内重组FDH实时表达的一种非入侵方法。

关键词:

English

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

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

Corresponding author: 黄庶识,陈丽梅; 黄庶识,陈丽梅

Received Date: 05 June 2012
Fund Project:
本文系国家自然科学基金(No.30970263)资助项目

Abstract: 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.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

单细胞激光拉曼光谱检测重组大肠杆菌细胞表达甲酸脱氢酶

通讯作者: 黄庶识,陈丽梅; 黄庶识,陈丽梅

English

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

Corresponding author: 黄庶识,陈丽梅; 黄庶识,陈丽梅

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

单细胞激光拉曼光谱检测重组大肠杆菌细胞表达甲酸脱氢酶

通讯作者: 黄庶识,陈丽梅; 黄庶识,陈丽梅

English

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

Corresponding author: 黄庶识,陈丽梅; 黄庶识,陈丽梅

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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