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Pre- study isothiocyanate fluorescein - the preparation of lysozyme and its impact on Lysozyme Crystal Growth

Author: DaiLianHua
Tutor: LiuFengLing
School: Shandong Normal University
Course: Organic Chemistry
Keywords: protein crystal growth lysozyme Fluorescein-5-Lsothiocyanalte-protein confocal laser scanning microscopy
CLC: O629
Type: Master's thesis
Year: 2005
Downloads: 113
Quote: 0
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Abstract


In the study of the macromolecules, the basic knowledge that we want to know is the process of the protein growth, the physical chemistry of the protein growth and how to receive the good quality protein crystal. The study of the process of the protein crystal growth is increasingly paid attention to because the protein crystal growth has play an important role in the life sciences and the microgravity sciences. In the paper, we introduced the significance for the protein crystal growth and the useful tools and the methods for the studying various aspects of protein crystal growth, such as static and dynamic lighting scattering, atomic force microscopy and the laser confocal microscopy and so on. Meanwhile we simply introduced some methods of protein crystallization. It is well known that the fluorescence is one of the most versatile and powerful tool for the studying of macromolecules, while most proteins are intrinsically fluorescent, working at crystallization concentrations requires the use of covalently prepared derivatives added as tracers. This approach requires derivatives that do not markedly affect the crystal packing. We have prepared fluorescent derivatives of lysozyme. Fluorescein-5-Lsothiocyanalte (FITC) was used as a probe to label the lysozyme. The absorption fluorescence spectra of the labeled lysozyme indicate that FITC has little effect on the biological and chemical characteristics of lysozyme. So the FITC derivatives of lysozyme can be used in research of protein crystal growth by the laser scanning confocal microscopy.

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