苋菜amaAG基因克隆与生物信息学分析

Cloning and Bioinformatics Analysis of ama AG in Amaranthus tricolor L.

以苋菜种子为初始材料,通过离体培养获得苋菜各阶段试管苗,利用实验室构建的苋菜开花过程转录组数据库(NCBI SRA number:SRR924089,SSR924090,SRR924091,SRR924092),采用RT-PCR结合RACE技术进行ama AG基因全长c DNA序列克隆,然后对其进行生物信息学分析。结果表明,苋菜ama AG基因全长为1 123 bp,其中5'UTR 122 bp,3'UTR 243 bp,poly A尾长17 bp,并包含一个741 bp的开放阅读框,编码246个氨基酸。通过生物信息学分析可知,该蛋白属于亲水蛋白,但不是稳定蛋白;属于MADS和K-box超级家族,包含MADS_MEF2-like和K-box功能位点,与植物开花相关。ama AG基因差异表达分析表明,该基因是在开花期表达量最高。说明本研究已获得了苋菜ama AG基因,这为将来深入研究其功能奠定了基础,也为研究苋菜开花分子机制提供参考。

The seeds were used to culture in vitro plantlets of Amaranthus tricolor as the material. The full-length ama AG c DNA sequence was cloned using transcriptome database of A. tricolor（ NCBI SRA number： SRR924089,SSR924090,SRR924091,SRR924092）,RT-P CR and RACE techniques,and the bioinformatics was analyzed. The full-length ama AG c DNA sequence was 1 123 bp in length,containing an open reading frame of 741 bp（ encoding 246 animo acids）,flanked by 122 bp 5 ＇ UTR and 243 bp 3 ＇UTR that contained a 17 bp poly A. Bioinformatics analysis showed that the protein was hydrophilic cytoplasmic protein,but not stable protein. The protein belongs to MADS and K-box superfamily with MADS_MEF2-like and K-box functional sites and the two sites were related to the growth of plants. The analysis of ama AG gene differential expression indicated that the gene expression level was the highest during flowering,which showed that the gene was cloned. The study is helpful for further research of ama AG function and provides references for flower molecular mechanism of Amaranthus tricolor.