小单孢菌3′,4′-双脱羟基酶基因功能的研究
Function of 3′,4′ -double dehydroxylase gene from Micromonospora
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摘要: 根据文献查阅、同源比对及进化树分析，推测绛红色小单孢菌中gntI和伊纽小单孢菌sisI为3′,4′-双脱羟基酶基因。通过克隆得到gntI和sisI，随后将其构建到pET30a表达载体，并在E.coli BL21实现异源表达。利用生物信息学对该蛋白亲水性图谱、空间结构及活性域进行分析、预测。基于同源建模得到三级结构，推测其二级结构以α-螺旋为主，肽段40～60,100～120处可能为该酶活性中心。此研究为该酶进一步功能研究和应用奠定了基础。
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关键词:
- 小单孢菌 /
- 3′,4′-双脱羟基酶基因 /
- 生物信息学
Abstract: Through research in literature,homologous alignment and phylogenetic analysis,gntI from Micro- monospora purpurea and sisI from Micromonospora inyoensis were presumed to be 3′,4′ -double dehydroxylase gene.Then the nucleic acid sequence of gntI and sisI were amplified and ligated to expression vector pET30 for heterologous expression in E.coli BL21. Bioinformatics was utilized to analyze the hydropathy plot ，structure and catalytic domain of the protein GntI and sisI. Based on homologous modeling,the spatial structures were gained,speculating that α-helix was the main secondary structure,and catalytic domain may be located at 40-60,100-120 amino acids.This study established the foundation for further functional research and potential application of 3′,4′ -double dehydroxylase.-
Keywords:
- Micromonospora /
- 3′,4′ -double dehydroxylase /
- bioinformatics
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