拟南芥4-香豆酰-CoA连接酶基因的克隆与生物信息学分析

Cloning and Sequence Analysis of a 4-coumaryl: CoA ligase Gene from Arabidopsis thaliana

以拟南芥总RNA为模板,利用RT-PCR的方法从拟南芥中扩增获得一条4-香豆酰-CoA连接酶基因完整的cDNA序列,命名为4CL1。利用生物信息学生物软件对其核酸和蛋白质序列进行分析,结果表明,该序列长1 686bp,与已报道的拟南芥4-香豆酰-CoA连接酶基因的序列相似性达到98%~100%,氨基酸序列相似性为99%~100%;4CL1基因编码561个氨基酸,该氨基酸序列具有典型的4-香豆酰-CoA连接酶AMP-binding保守结构域LPYSSGTTGLPK,同时含有该蛋白家族的催化活性中心GEICIRG序列;预测的分子量为61.05kDa,理论等电点为5.25,不稳定参数为35.93,在分类上属于稳定性蛋白;二级结构主要以无规则蜷曲、α-螺旋、延伸链及β转角为主,其含量分别为34.22%、30.30%、24.06%和11.41%。

A complete cDNA sequence of 4-coumaryl:CoA ligase1 gene was obtained from Arabidopsis thaliana total RNA by RT-PCR method, named 4CL1. The results of multiple sequence alignment showed that 4CL1 was of 1 686 bp with 98% ~ 100% nucleic acid sequence similarity and 99% ~ 100% amino acid sequence similarity to the reported Arabidopsis thaliana 4-coumaryl: CoA ligase 1. Domain analysis results revealed that the AMP-binding characteristics sequences of LPYSSGTTGLPKGVLFGPGLT and the active site sequences GEICIRG of 4-coumaryl:CoA ligase protein family were found in cloned 4CL1 amino acids sequence. The predicted molecular weight was 61.05 kDa and the theoretical isoelectric point was 5.25. The calculated instability parameter was 35.93, which indicated that this protein was stable in classification. The secondary structure of resveratrol synthase mainly contained random curling, α-helix, extended strand and β-turn, which content was 34.22%, 30.30%,24.06% and 17.66%, respectively.