枯草芽孢杆菌α-乙酰乳酸脱羧酶基因(alsD)的克隆和生物信息学分析

doi:10.11924/j.issn.1000-6850.casb18020058

中国农学通报 ›› 2019, Vol. 35 ›› Issue (21): 96-102.doi: 10.11924/j.issn.1000-6850.casb18020058

所属专题：生物技术

枯草芽孢杆菌α-乙酰乳酸脱羧酶基因(alsD)的克隆和生物信息学分析

刘文娟, 黄守锋, 葛菁萍

₁黑龙江大学生命科学学院

收稿日期:2018-02-12 修回日期:2018-04-13 接受日期:2018-04-20 出版日期:2019-08-13 发布日期:2019-08-13
通讯作者: 葛菁萍
基金资助:
国家自然科学基金“从2,3-丁二醇代谢角度构建工程微生物群体及其生态学机制研究”(31570492)；黑龙江省高等学校科技创新团队“农业微生物发酵技术”(2012td009)。

Cloning and Bioinformatical Analysis of α-acetolactate Decarboxylase Gene (alsD) in Bacillus subtilis

Received:2018-02-12 Revised:2018-04-13 Accepted:2018-04-20 Online:2019-08-13 Published:2019-08-13

摘要/Abstract

摘要： [目的]α-乙酰乳酸脱羧酶是2,3-丁二醇生物合成途径的关键酶之一,利用生物信息学方法对α-乙酰乳酸脱羧酶基因(alsD)进行分析。[方法]根据GenBank中α-乙酰乳酸脱羧酶基因(alsD)的基因序列设计引物,以枯草芽孢杆菌(Bacillus subtilis)CICC10026基因组DNA为模板,通过PCR扩增得到Bacillus subtilis CICC10026基因片段alsD。对该片段进行序列测定,利用生物信息学分析软件对该序列进行同源性分析、二级结构和功能性分析及3D 结构预测等。[结果] 结果表明,获得的alsD片段为768bp,未发生碱基突变,该片段为枯草芽孢杆菌(Bacillus subtilis)CICC10026的α-乙酰乳酸脱羧酶基因(alsD)的基因序列。[结论]本研究结果为构建产2,3-丁二醇的工程菌株奠定了理论基础。

关键词: 疏勒河中游, 疏勒河中游, 绿洲, 生态功能分区, 生态环境

Abstract: α-acetolactate decarboxylase is one of the key enzymes in 2,3-butanediol biosynthesis pathway, and the alsD gene is analyzed by bioinformatics method. Primers are designed based on the gene sequence of α- acetolactate decarboxylase gene (alsD) in GenBank. Bacillus subtilis CICC10026 gene fragment alsD is amplified by PCR which the template is Bacillus subtilis CICC10026 genomic DNA. The fragment is sequenced and analyzed by bioinformatics software for homology analysis, secondary structure and functional analysis and 3D structure prediction. The results showed that the obtained alsD fragment was 768 bp without base mutation, and the fragment was the alsD gene sequence of Bacillus subtilis CICC10026. The results of this study lay a foundation for the construction of engineering strains which could produce 2,3-butanediol.

刘文娟,黄守锋,葛菁萍. 枯草芽孢杆菌α-乙酰乳酸脱羧酶基因(alsD)的克隆和生物信息学分析[J]. 中国农学通报, 2019, 35(21): 96-102.

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枯草芽孢杆菌α-乙酰乳酸脱羧酶基因(alsD)的克隆和生物信息学分析

Cloning and Bioinformatical Analysis of α-acetolactate Decarboxylase Gene (alsD) in Bacillus subtilis

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