Study of Fluorescence Spectra of Blattela Germanica Allergen, Derenatured and Renatured Bla g 2
ZHU Jian-qi1, LIU Zhi-gang1*, HUANG Hai-zhen2, ZHU Yong-feng1, XU Hong1
1. College of Life Science, Shenzhen University, Shenzhen 518060, China 2. Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China
摘要： 包涵体中的重组蛋白经抽提后可以在变性状态下纯化，而纯化后的复性过程是基因工程下游处理的重要环节。通过对变应原Bla g 2蛋白复性前后荧光光谱的比较和分析、变性剂(尿素和SDS)对复性后Bla g 2蛋白的荧光滴定实验、以及复性后Bla g 2在不同pH下的荧光光谱分析，推断出Bla g 2蛋白分子在不同环境下构象的变化及其光谱学特征，初步建立了一种新的检测重组变应原蛋白变复性的光谱实验方法。
关键词：德国小蠊;变应原;Bla g 2;变性;复性;荧光光谱
Abstract：Recombinant proteins extracted from inclusion body remain in denaturation status. Renaturation in vitro after initial purification is a key step of downstream processing. A common method of renaturation of recombinant proteins is the dilution method. With Bla g 2 as a model protein, the conformational changes of denatured and renatured Bla g 2 were investigated by applying fluorescence spectra. The effects of different urea concentrations, different SDS concentrations and different pH on the fluorescence intensity of renatured protein were also investigated. The reasons for these were studied with the knowledge of molecular structure.
Key words：Blattella germanica;Allergen;Bla g 2;Denaturation;Renaturation;Fluorescence spectra
朱健琦1,刘志刚1*,黄海珍2,朱永烽1,徐宏1 . 德国小蠊变应原Bla g 2蛋白变复性的荧光光谱研究[J]. 光谱学与光谱分析, 2006, 26(12): 2254-2257.
ZHU Jian-qi1, LIU Zhi-gang1*, HUANG Hai-zhen2, ZHU Yong-feng1, XU Hong1 . Study of Fluorescence Spectra of Blattela Germanica Allergen, Derenatured and Renatured Bla g 2. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2006, 26(12): 2254-2257.
[1] Alla G, Mi L, Sabina W. J. Mol. Biol., 2005, 348: 433. [2] Jaenicke R. In:Creighton T E. ed. Protein Structrue: A Practical Approach, IRL, 1989. [3] XU Ming-bo, MENG Wen-hua, MA Xian-kai(徐明波, 孟文华，马贤凯). Chinese Biochemical Journal(生物化学杂志), 1994, 10(3): 376. [4] FU Ya-zhen(傅亚珍). Acta Biophysica Sinica(生物物理学报), 1995, 11(3): 295. [5] YAN Yuan, XU Jin-gou, CHEN Guo-zhen(鄢 远, 许金钩, 陈国珍). Science in China(中国科学), 1997, 27(1): 16. [6] CHEN Guo-zhen，HUANG Xian-zhi, ZHENG Zhu-zi, et al(陈国珍, 黄贤智，郑朱梓，等). Fluorescent Analytical Method(荧光分析法). Beijing: Science Press(北京: 科学出版社), 1990. 131. [7] WEI Xiao-fang, DING Xi-ming, LIU Hui-zhou(魏晓芳, 丁西明, 刘会洲). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2000, 20(4): 556. [8] ZHANG Jing(张 静). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2004, 24(4): 455. [9] Burstein E A, Vedenkina N S, Ivkova M N. Photobio.,1973, 8: 267. [10] Guo Q, Zhao F, Guo Z. J. Biochem., 2004, 136(1): 49. [11] WANG Fan, ZHOU Hai-meng(王 帆，周海梦). Progress in Natural Science(自然科学进展)，1995, 5(2): 216. [12] ZHOU Cheng-lu(邹承鲁). Second Genetic Code-Study of New Synthesis Peptide and Protein Folding(第二遗传密码—新生肽链及蛋白质折叠研究). Changsha: Hunan Science and Technology Press(长沙: 湖南科学技术出版社), 1997. 51.
