草海桐铜锌超氧化物歧化酶基因的克隆及序列分析  

Cloning and sequence analysis of copper/zinc-superoxide dismutase gene from Scaevola sericea

作　　者：张静文 张力文 严云香 常义 陈艳 李欣勇 ZHANG Jing-wen;ZHANG Li-wen;YAN Yun-xiang;CHANG Yi;CHEN Yan;LI Xin-yong(College of Life Sciences,Hainan Normal University/Ministry of Education Key Laboratory for Ecology of Tropical Islands,Haikou 571158,China;Institute of Tropical Crop Genetic Resources,Chinese Academy of Tropical Agriculture Sciences,Danzhou,Hainan 571737,China)

机构地区：[1]海南师范大学生命科学学院/热带岛屿生态学教育部重点实验室,海口571158 [2]中国热带农业科学院热带作物品种资源研究所,海南儋州571737

出　　处：《西南农业学报》2022年第2期425-431,共7页Southwest China Journal of Agricultural Sciences

基　　金：海南省自然科学基金(2019RC181);海南师范大学博士教授科研启动项目(301060181825)。

摘　　要：【目的】铜锌超氧化物歧化酶(Cu/Zn-SOD)在保护细胞和组织免受氧化损伤方面起重要作用。对滨海植物草海桐盐胁迫诱导的Cu/Zn-SOD进行克隆和序列分析,为揭示草海桐适应盐碱环境提供理论依据。【方法】以盐胁迫的草海桐为材料,运用RACE技术克隆草海桐Cu/Zn-SOD,命名为SsCSD,采用生物信息学方法进行序列分析,构建pBinGlyRed-SsCSD重组质粒。【结果】通过RACE技术获得SsCSD基因全长共1148 bp,其最大开放阅读框(ORF)为687 bp,编码228个氨基酸。氨基酸保守结构域分析表明,该序列具有铜锌超氧化物歧化酶保守结构域,属于SOD家族。生物信息学分析表明其定位于叶绿体中,氨基酸序列分析显示其与已知植物叶绿体Cu/Zn-SOD的氨基酸残基高度同源。同时将SsCSD基因与植物表达载体pBinGlyRed进行重组,并成功转入农杆菌GV3101中。【结论】草海桐铜锌超氧化物歧化酶氨基酸序列与同源物种相似度高,在进化上高度保守,对进一步通过转基因验证该酶生物学功能具有重要意义。[Objective]Copper zinc superoxide dismutase(Cu/Zn-SOD)played an important role in protecting cells and tissues from oxidative damage.In the present study,Cu/Zn-SOD were cloned and sequence analyzed from salt-stressed Scaevola sericea.The findings will provide a scientific basis for further revealing the adaptation mechanism of S.sericea to the saline-alkali environment.[Method]The cDNA sequence of Cu/Zn-SOD was obtained by RACE,named as SsCSD.The bioinformatics analysis was performed to research the SsCSD,and the recombinant plasmid pBinGlyRed-SsCSD was constructed.[Result]The full length of SsCSD was 1148 bp and contained an open reading frame of 687 bp,which encoded 288 amino acid residues.Amino acid sequence analysis revealed that there was a typical Cu/Zn-SOD functional domain,belonging to SOD family.The bioinformatics analysis showed that it was located in the chloroplast.Meanwhile,evolution and clustering analysis showed that the Cu/Zn-SOD protein of S.sericea was highly homologous with known plant of Cu/Zn-SOD protein in chloroplasts.Besides,the recombinant plasmid pBinGlyRed-SsCSD was constructed and transferred to Agrobacterium tumefaciens GV3101.[Conclusion]We obtained the full length of SsCSD,which the amino acid sequence was highly similar with the known species and highly conserved in evolution,which was of great significance to further reveal the biological function of the Cu/Zn-SOD in S.sericea.

关 键 词：草海桐 Cu/Zn-SOD 基因克隆 序列分析 

分 类 号：Q943.2[生物学—植物学]