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

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。

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

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.