牡丹长链脂酰辅酶A合成酶基因PsLACS的克隆及生物信息学分析

Molecular Cloning and Bioinformatics Analysis of PsLACS Gene from Tree Peony(Paeonia suffruticosa)

根据本实验室前期获得的长链脂酰辅酶A基因的EST序列设计引物,本研究利用RACE扩增从牡丹中得到2 377 bp的Ps LACS全长c DNA,其中,包括5'UTR 347 bp,3'UTR 200 bp和1 830 bp的编码区,共编码609个氨基酸,分子量为67.94 k D。Ps LACS包括30个可能的磷酸化位点,其中14个位于丝氨酸(S),7个位于苏氨酸(T),9个位于酪氨酸(Y),这可能与酶活性的调节有关。二级结构预测显示,Ps LACS含α-螺旋和无规则卷曲较多。同源性分析结果表明,Ps LACS与葡萄LACS同源性最高,其次是橙子LACS、蓖麻LACS。系统进化树分析结果与同源性比对的结果基本一致。研究结果可为进一步研究牡丹Ps LACS基因的功能提供了理论基础。

Based on the EST of Long-chain Acyl-coenzyme A(LACS) from our laboratory, the full length c DNA was obtained from Tree peony(Paeonia suffruticosa Andr.) by RACE amplification and named as Ps LACS. The length c DNA of Ps LACS was 2 377 bp including 5'UTR of 347 bp, 3'UTR of 200 bp and 1 830 bp encoding frame,which encoded 609 amino acids and had a molecular weight of 67.94 k D. The predication of phosphorylation sites showed that the amino acid sequence of Ps LACS had 30 possible phosphorylation sites including 14 Serine sites, 7Threonine sites and 9 Tyrosine sites, which might be associated with the regulation of enzyme activity. The secondary structures of Ps LACS were mainly α-helix and random coil. Alignment analysis was consistent with phylogenetic analysis, which indicated that Ps LACS should be highly conserved with that of Vitis vinifera LACS,Citrus sinensis LACS, and Ricinus communis LACS, and basically accorded with botanical classification system.All the results were the necessary materials for further study of the function of Ps LACS gene in Tree peony.