Cloning and Bioinformatic Analysis of Potato α-Amylase Gene amyA1
In this research, we amplified a cDNA for potato α-amylase by RT-PCR and cloned it. Sequence analysis showed that the cDNA had a 1 224 bp open reading frame and was referred to as amyA1, which encodes for an α-amylase with 407 amino acid residues (GenBank accession number: GQ406048.1) with the MW 46.40 kD.After that we used semi-quantitative RT-PCR assay to detective the expression of the amyA1 gene in potato leaves and stems. The result showed that the expression in stems is a little stronger than in leaves. Then we analyzed the amino acid sequence bioinformatically, including its codon usage bias, physical and chemical properties, subcellular localization, and conserved structures. 29 α-amylase genes from same or different species were taken from the GenBank for constructing a phylogenetic tree. The bioinformatical analyses showed that the putative protein shared 98% identity with a published potato α-amylase (GenBank accession number: M79328.1) at the amino acid level. The deducted α-amylase also contains a catalytic domain (PF00128、 SM00624) between 20 to 348 and a C-terminal beta-sheet domain (PF07821)between 349～407, which are similar to ones of the amylase family 13.The postulated eight-stranded alpha/beta barrel was also found in the enzyme, which was thought as an active site of α-amylase. According to the phylogenetic tree, the two genes from potato presents more close homology to those from cassava and apple than from barley, rice and maize.
Potatoα-amylaseCloneBioinformatic analysis
王玉丽、王永刚、马建忠、王倩、陶鹏飞、苏移
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兰州理工大学生命科学与工程学院,兰州,730050
马铃薯 α-淀粉酶基因 克隆 生物信息学
Technical Bureau, GansuDegree Courses on Building Lan Zhou University of Technology
NETL
NSTL
万方数据
维普
