东亚砂藓Cu/Zn SOD基因的克隆及序列分析

目的]对东亚砂藓(Rhacomitrunm japonicum)的Cu/Zn SOD基因进行克隆,并对其序列进行分析。方法]利用RT-PCR技术获得东亚砂藓Cu/Zn SOD基因的cDNA,同时对该序列进行生物信息学分析。结果]东亚砂藓Cu/Zn SOD基因cDNA长565 bp,包含一个长为465 bp的ORF,编码154个氨基酸残基,预测的该蛋白的分子量为15.5 kD,理论等电点为5.77,负电荷残基(Asp+Glu)总数为18个,正电荷残基(Arg+Lys)总数为12个,不稳定系数是13.36;其编码的氨基酸序列包含了5个蛋白保守区,均为铜锌超氧化物歧化酶超家族所有;Blastn比对表明东亚砂藓Cu/Zn SOD基因与毛尖紫萼藓、小立碗藓相关基因的同源性高达99%和82%。结论]该研究为进一步研究紫萼藓科植物的抗旱性及苔藓Cu/Zn-SOD在抗非生物胁迫方面的功能提供了理论依据与基础。

Cloning and Sequence Analysis of Cu/Zn SOD from Rhacomitrunm japonicum

Objective] The aim was to clone and analyze the sequence of Cu/Zn SOD cDNA from Rhacomitrunm japonicum.Method] The cDNA of Rhacomitrunm japonicum Cu/Zn SOD was cloned by RT-PCR and then its sequence was analyzed by bioinformatics method.Result] The cDNA sequence of Rhacomitrunm japonicum Cu/Zn SOD was 565 bp,containing a 465 bp ORF,encoding 154 amino acids,which prediction molecular weight and theoretical isoelectric point of was 15.5 kD and 5.77;the protein had 18 negative charge residues(Asp and Glu) and 12 positive charge residues(Arg and Lys),unstable coefficient was 13.36;the sequence of amino acids encoded by Cu/Zn SOD gene contains five protein conserved regions and all of them belonged to Cu/Zn superoxide dismutase superfamily;the Blastn comparison analysis showed that the homology between Rhacomitrunm japonicum and Grimmia pilifera,Physcomitrella patens was 99% and 82%.Conclusion] This study provided theory basis and foundation for the further research of drought resistance of Grimmiaceae plants and function of moss Cu/Zn-SOD in the abiotic stresses resistance.