广藿香α-淀粉酶PcAMY1基因克隆和序列分析

Cloning and Analysis of Alpha-amylase(PcAMY1) Gene in Pogostmen cablin

为了解广藿香对淀粉的利用,克隆了广藿香(Pogostemon cablin)的α-淀粉酶(alpha-amylase,AMY)基因,对其进行相关生物信息学分析。搜索本课题组建立的广藿香转录组数据库,获得AMY基因全长序列,并设计全长引物进行PCR验证;利用生物信息学软件对AMY基因进行生物信息学分析。本研究获得的广藿香AMY基因命名为PcAMY1基因(GenBank登录号为KC862311),该基因全长1 420 bp,编码422个氨基酸。基于生物信息软件分析了PcAMY1基因编码蛋白的理化特性。系统进化树分析结果表明,PcAMY1基因与牵牛(Ipomoea nil)的AMY序列同源性最高,与赤豆(Vigna angularis)、菜豆(haseolus vulgaris)、蒺藜苜蓿(Medicago truncatula)等植物次之,与自然进化关系保持一致。PcAMY1基因在广藿香嫩茎、成熟茎、老茎、嫩叶、成熟叶、老叶中均有表达,但在老茎中表达量最高。成功克隆到广藿香PcAMY1基因,并对其进行相关生物信息学与基因表达分析。

Objective In order to understand the using process of starch in Pogostemon ceblin, an alpha-amylase gene PcAMY1 was cloned and analyzed bioinformatically. Methods The full length AMY gene sequence was retrieved from transcriptomic database of P. cablin, the full length primer was designed for PCR verification, and PcAMY1 gene was analyzed using several bioinformatical softwares. Results PcAMY1 （NCBI accession, KC862311） containedl 420 bp length of open reading frame （ORF） and encoded 422 amino acids postulated. Additionally, the physical and chemical properties of PcAMYl-coded protein were analyzed by bioinformatical softwares. Phylogenetic analysis showed that PcAMY1 was tightly clustered with AMY genes in Ipomoea nil, Vigna angularis, and haseolus vulgaris, which is consistent to the phylogenetic relationship of species. The expression level of PcAMY1 was most abundantly in P. cablin old stem, followed by mature leaf, that for other organs was low. Conclusion PcAMY1 is successfully coloned and molecularly characterized. The results would lay a fundamental foundation for P. cablin researches.