小麦FeSOD基因的克隆与序列分析

Cloning and Sequence Analysis of Stress Tolerance Relative Gene FeSOD in wheat(Triticum aestivum)

超氧化物歧化酶(superoxide dismutase,SOD)是一种广泛存在于真核生物中的金属酶类,在植物的抗逆性中起到重要的作用。本文以小麦烟农19号为材料,提取植株叶片总RNA,根据NCBI公布的FeSOD基因序列设计简并引物,采用RT-PCR和RACE技术首次获得小麦抗逆基因FeSOD(GenBank:JX398977)的全长cDNA序列。生物学软件分析表明,该序列全长为1271bp,5'UTR为187bp,3'UTR为490bp,开放阅读框长度为591 bp,编码197个氨基酸,分子质量为22.8KD,等电点(PI)为5.382,经BlastP和Clustalx进行比对发现,小麦FeSOD的氨基酸序列与玉米、水稻等物种具有高的同源性;同时,利用Mega5软件进行系统谱系分析,发现小麦FeSOD与玉米和水稻的亲缘关系最近。

Superoxide dismutase（SOD） is an important enzyme which exists in eukaryote extensively and plays an essential role in stress-tolerance of higher plants.In this study,the total RNA was extracted from leaves of Yannong19,and the degenerate primers were designed according to the conserved regions of FeSOD gene sequence in Genbank.The full-length cDNA sequence of FeSOD in wheat（GenBank：JX398977） was obtained by RT-PCR and RACE technology.Biology software analysis showed that the full-length of the sequence was 1271bp with a 187bp 5＇UTR,a 490bp 3＇UTR,and a 591bp open reading frame encoding 197 amino acids.The molecular weight was 22.8KD,and its isoelectric point（PI） was 5.382.The sequence with FeSOD of other plants was blasted on NCBI website,and it was found that the FeSOD amino acids sequence in wheat had high homology with other plants（maize and rice and other species）.At the same time,we found the genetic relationship of FeSOD in wheat was the most close to that of corn and rice by Mega5 software analysis.