编码酿酒酵母丙酮酸脱羧酶(Pdc6)基因克隆及其生物信息学分析

doi:10.11924/j.issn.1000-6850.casb2020-0296

中国农学通报 ›› 2021, Vol. 37 ›› Issue (9): 103-108.doi: 10.11924/j.issn.1000-6850.casb2020-0296

所属专题：生物技术

编码酿酒酵母丙酮酸脱羧酶(Pdc6)基因克隆及其生物信息学分析

王长丽¹^,²(), 廖巍¹, 叶广彬¹, 葛菁萍²^,³, 刘磊², 马毓坚¹, 黄霞¹, 宾晓芸¹()

¹右江民族医学院,广西百色 533000
²黑龙江大学生命科学学院微生物省高校重点实验室,哈尔滨 150080
³农业微生物技术教育部工程研究中心,哈尔滨 150500

收稿日期:2020-07-27 修回日期:2020-10-19 出版日期:2021-03-25 发布日期:2021-04-09
通讯作者: 宾晓芸
作者简介:王长丽,女,1992年出生,黑龙江肇东人,助教,硕士,研究方向：微生物资源挖掘与利用。通信地址：533000 广西百色市右江区城乡路98号校团委,Tel：0776-2835595,E-mail： 1808907124@qq.com。
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究”(31570492);广西自然科学基金“广西仫佬族人群骨密度与瘦素受体基因多态性的相关性研究”(2017JJA10377);广西高校中青年教师基础能力提升项目“乳酸菌胞外多糖的结构表征及其抗肿瘤活性研究”(2020KY13007);右江民族医学院2019年度校级科研课题“鸡黏膜免疫中CpG ODN最优类型和剂量的确定及其机制初探”(yy2019ky013)

Pyruvate Decarboxylase (Pdc6) Gene Cloning and Bioinformatics Analysis in Saccharomyces cerevisiae

Wang Changli¹^,²(), Liao Wei¹, Ye Guangbin¹, Ge Jingping²^,³, Liu Lei², Ma Yujian¹, Huang Xia¹, Bin Xiaoyun¹()

¹Youjiang Medical University for Nationalities, Baise Guangxi 533000
²Key Laboratory of Microbiology, College of Heilongjiang Province, School of Life Sciences, Heilongjiang University, Harbin 150080
³Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education, Harbin 150500

Received:2020-07-27 Revised:2020-10-19 Online:2021-03-25 Published:2021-04-09
Contact: Bin Xiaoyun

摘要/Abstract

摘要：

旨在从生物信息学角度更好地研究酿酒酵母丙酮酸脱羧酶(Pdc6)的结构和功能,克隆酿酒酵母H5的丙酮酸脱羧酶基因pdc6。利用Primer 5.0软件设计1对20 bp引物,以H5基因组DNA为模板克隆获得pdc6,并利用生物信息学分析方法对其序列进行验证及分析。该序列长度为1692 bp,无碱基缺失,且该基因具有完整的开放阅读框,该基因编码的Pdc6包含563个氨基酸残基,该蛋白质中酸性氨基酸残基数量和碱性氨基酸残基数量大致相同;Pdc6理论等电点5.8,疏水性最大值为2.32,亚细胞定位预测其位于细胞外基质,属于酸性蛋白质,并预测了空间结构模型。本研究说明该蛋白结构与Pdc1和Pdc5结构类似,对酿酒酵母H5编码丙酮酸脱羧酶(Pdc6)基因序列克隆和生物信息学分析后,可为后续单基因敲除选取基因顺序的研究提供一定的理论基础。

关键词: 酿酒酵母, 生物信息学分析, 克隆, 丙酮酸脱羧酶基因, 开放阅读框

Abstract:

The aims are to study the structure and function of pyruvate decarboxylase (Pdc6) in Saccharomyces cerevisiae, and clone pdc6 gene in S. cerevisiae H5. A pair of 20 bp primers was designed by Primer5.0 software. The pdc6 gene was cloned using the genomic DNA of S. cerevisiae H5 as template, and the cloned sequence was subsequently verified and analyzed by bioinformatics method. The results show that the sequence length is 1692 bp with a completely open reading frame (ORF) without deletion. Besides, the Pdc6 protein contains 563 amino acid residues and the number of acidic amino acid is almost the same as that of the basic amino acid residues. The theoretical isoelectric point of Pdc6 is 5.8 and the maximum hydrophobicity is 2.32. Subcellular localization predicts that it is located in extracellular matrix and belongs to acidic protein, and the spatial structure model is predicted. This study shows that the structure of Pdc6 protein is similar to the Pdc1 protein and the Pdc5 protein. The bioinformatically analysis of the gene sequence of pyruvate decarboxylase (Pdc6) encoded by S. cerevisiae H5 can provide a theoretical basis for the subsequent study of gene sequence selection for single gene knockout.

Key words: Saccharomyces cerevisiae, bioinformatics analysis, clone, pyruvate decarboxylase (Pdc6) gene, ORF

中图分类号:

S216.2

王长丽, 廖巍, 叶广彬, 葛菁萍, 刘磊, 马毓坚, 黄霞, 宾晓芸. 编码酿酒酵母丙酮酸脱羧酶(Pdc6)基因克隆及其生物信息学分析[J]. 中国农学通报, 2021, 37(9): 103-108.

Wang Changli, Liao Wei, Ye Guangbin, Ge Jingping, Liu Lei, Ma Yujian, Huang Xia, Bin Xiaoyun. Pyruvate Decarboxylase (Pdc6) Gene Cloning and Bioinformatics Analysis in Saccharomyces cerevisiae[J]. Chinese Agricultural Science Bulletin, 2021, 37(9): 103-108.

图/表 2

参考文献 27

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编码酿酒酵母丙酮酸脱羧酶(Pdc6)基因克隆及其生物信息学分析

Pyruvate Decarboxylase (Pdc6) Gene Cloning and Bioinformatics Analysis in Saccharomyces cerevisiae

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