花生FAD8基因的克隆及生物信息学分析

Cloning and bioinformatics analysis of FAD8 gene from peanut( Arachis hypogaea)

[目的]克隆重要油料作物花生中在多不饱和脂肪酸合成代谢途径发挥重要作用的脂肪酸脱氢酶FAD8基因,并对其进行生物信息学分析。[方法]以高油花生品种鲁花14为材料,电子拼接花生FAD相关EST序列并结合RT-PCR方法克隆花生Ah FAD8全长c DNA,并对其基因序列特征进行分析。[结果]花生Ah FAD8基因c DNA全长1 494 bp,开放阅读框为1 359 bp,编码453个氨基酸,分子量为51.71 k Da;蛋白序列比较发现花生FAD8与野大豆和普通大豆的相似性最高,为83%;NJ法聚类分析表明其与上述两种物种的亲缘关系最近,且支持率达96%。[结论]从花生中分离到一个新的ω-3脂肪酸脱氢酶基因,具有FAD8蛋白的保守结构域及3个富组氨酸基序,属于FAD8家族。

[ Objective] Cloning and bioinformatics analysis of fatty acid desaturases gene FAD8 which is a key enzyme gene in the formation of unsaturated fatty acids in the important oil crop of peanut. [ Methods] Basis on the electronic splicing of ESTs of FAD from peanut and with the RT - PCR method,the high oil peanut species Luhua 14 was used as research material to iso- late the full length cDNA of AhFAD8, then sequence analysis of FAD8 were performed with the bioinformatics. [ Results ] The results showed that, the cDNA of AhFAD8 gene was 1 494 bp,including an ORF of 1 359 bp which contained 453 amino acids and the molecular mass of AhFAD8 was 51.71 kDa. Protein sequence alignments showed that FAD8 of peanut had the highest identity 83% with Glycine soja and Glycine max. The phylogenetic tree analysis using NJ method showed that it clustered closely with 96% bootstrap support with FAD family of Glycine soja and Glycine max. [ Conclusion] A new co -3 fatty acid desaturase gene was isolated from peanut,which contained the conserved domains of FAD8 proteins and three histidine -rich motifs, and it belonged to FAD8 family.