河北师范大学学报—

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刊名：河北师范大学学报（自然科学版）Journal of Hebei Normal University (Natural Science)
主办：河北师范大学
ISSN： 1000-5854
CN： 13-1061/N
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SLA-Ⅰ重链分子的表达及结构分析

大连大学生物工程学院, 辽宁大连　116622
DOI：

Expression and Structure-analysis of SLA-Ⅰ Heavy Chain

GAO Feng-shan, FU Yin-kun

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摘要/Abstract

摘要：

为研究SLA-Ⅰ重链分子在pMAL-p2X载体表达条件,从培养基pH值、温度、菌体密度、IPTG浓度、时间5个条件设计了优化条件的实验.SDS-PAGE筛选出在pMAL-p2X系统上表达SLA-Ⅰ重链分子的最佳条件,并运用EXPASY服务器上的SOPMA法对SLA-Ⅰ重链分子和HLA-A2进行2级结构的预测,同时同源模建其3级结构.结果表明:pH8.0,37℃,0.2 mmol/LIPTG,培养5 h,D₆₀₀ nm=0.8是SLA-Ⅰ重链分子蛋白表达的最佳条件.2级结构预测和分析发现,猪和人MHCⅠ类分子重链2级结构成分上非常相似,其同源率远远大于氨基酸的同源率;同源模建发现,猪和人、小鼠的MHCⅠ类分子3级结构非常相似.

Abstract：

In order to detect the optimum expression condition for the SLA-Ⅰ heavy chain in pMAL-p2X expression system, the protein of interest was optimized from several aspects including different pH value of the medium, induction temperature, cell density, concent ration of IPTG and induction time. The interest of proteins w ere detected and analyzed by SDS-PAGE. Then by using SOPMA on EXPASY website, the secondary structure of SLA-Ⅰ heavy chain was predicted followed by homology modeling of SLA-Ⅰ heavy chain to predict its threedimensional structures (3D).The result s showed the optimal expression condition for the protein of interest was obtained (pH8. 0, 37 ℃,D₆₀₀ nm =0.8, 0.2 mmol/ LIPTG, 5 h).Prediction and analysis of the secondary structure of SLA-Ⅰ indicated that SLA-Ⅰ had similar secondary st ructure with human MHC class I and the homologous percentage of secondary st ructure elements between them w as much more than their homologous percentage of amino acids. Homology modeling of SLA-Ⅰ demonst rated SLA-Ⅰ had more similar 3D structure with human MHC Ⅰ than with mouse MHC.

关键词

关键词： SLA-Ⅰ重链分子 ; 表达条件 ; 2级结构 ; 3级结构

Key words： SLA-Ⅰ heavy chain ; expressing condition ; secondary structure ; 3D structure

参考文献 12

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