茅苍术乙酰辅酶A酰基转移酶基因(AlAACT)的克隆与序列分析

Cloning and Sequence Analysis of Acetyl-CoA Acetyltransferase Gene (AlAACT) from Atractylodes lancea

目的:对茅苍术乙酰辅酶A酰基转移酶基因(AlAACT)全长进行克隆,并分析其序列特征,为后期解析茅苍术萜类次生代谢产物的生物合成机制提供实验依据。方法:根据转录组测序所得的AACT基因片段设计引物,利用逆转录PCR技术获得全长cDNA序列,并采用生物信息学手段分析其序列特征。结果:克隆获得AlAACT基因的全长cDNA序列为1227 bp,编码408个氨基酸,理论相对分子质量为41.98 kDa,等电点为5.82,不存在跨膜区以及信号肽,为亲水性蛋白,具有多个磷酸化位点。进化树分析发现AlAACT基因与其他植物AACT基因具有较高相似性,尤其是菊科蓟属植物洋蓟,表明该基因保守性高。结论:成功获得茅苍术AlAACT基因序列,并掌握其序列特征,为后续深入研究该序列在茅苍术萜类化合物合成途径中的功能提供了必要的实验基础和理论依据。

Objective:To clone the full-length acetyl-CoA acyltransferase gene(AlAACT)from Atractylodes lancea and analyze its sequence characteristics,so as to provide experimental basis for later analysis of the biosynthesis mechanism of terpenoid secondary metabolites in Atractylodes lancea.Methods:The primers were designed based on the AACT gene fragments obtained from transcriptase sequencing,and the full-length cDNA sequence was obtained by reverse transcription PCR,the sequence characteristics were analyzed by bioinformatics.Results:The full-length cDNA sequence of AlAACT gene was 1227 bp,encoding 408 amino acids,with a theoretical molecular weight of 41.98 KDa and an isoelectric point of 5.82.It was a hydrophilic protein with multiple phosphorylation sites and no transmembrane region and signal peptide.Evolutionary tree analysis showed that AlAACT gene had high similarity with AACT gene of other plants,especially Cynara scolymus var.scolymus in Compositae,indicating that AlAACT gene was highly conserved.Conclusion:The AlAACT gene sequence of Atractylodes lancea is successfully obtained and its sequence characteristics are understood,it provides the necessary experimental basis and theoretical basis for further study of the function of this sequence in the terpenoid synthesis pathway of Atractylodes lancea.