水蕨(Ceratopteris thalictroides)查尔酮合成酶基因的克隆和表达(英文)

Isolation and Expression Profiling of Gene Encoding Chalcone Synthase in Ceratopteris thalictroides

查尔酮合酶(chalcone synthase,CHS)是植物类黄酮化合物合成的关键酶,有关蕨类植物CHS基因的序列及功能信息尚不完善。本研究采用快速扩增c DNA末端(RACE)技术克隆获得了模式蕨类植物——水蕨(Ceratopteris thalictroides)Ct CHS基因(Gen Bank登录号:JX027616.1),其c DNA序列全长为1616 bp,具有3个外显子和2个内含子,开放阅读框(ORF)为1215 bp,编码404个氨基酸。进化树分析表明,Ct CHS与问荆(Equisetum arvense)、松叶蕨(Psilotum nudum)和3种薄囊蕨的查尔酮合成酶基因聚为一枝,说明这些蕨类植物亲缘关系较近且为单系起源。通过构建原核表达体系成功获得Ct CHS蛋白的多克隆抗体并用于免疫印迹分析,结果表明Ct CHS基因的表达明显受紫外光(UV)诱导。Ct CHS基因的克隆与表达分析为进一步研究水蕨类黄酮化合物的合成及其调控机制提供了依据。

Chalcone synthase（ CHS） is a key enzyme in the synthesis of plant flavonoids.However,information on CHS genes in ferns is still unclear. In this study,rapid amplification of c DNA ends（ RACE） was used to isolate the full-length sequence of the CHS gene from model fern Ceratopteris thalictroides（ Ct CHS,Gen Bank accession number： JX027616.1）. Sequence analysis showed that the full length of the Ct CHS gene was 1616 bp,with three exons and two introns. Its ORF region was 1215 bp, encoding 404 amino acids. Phylogenetic analysis indicated that Ct CHS was clustered with the other ferns, including Equisetum arvense,Psilotum nudum and three leptosporangiate ferns,which reflected the monophyletic feature of the ferns according to Smith＇s system. Western blot analysis showed that the expression of this gene was significantly affected by ultraviolet（ UV） treatment. In this study,the full-length sequence of Ct CHS was cloned and the function of the Ct CHS protein was studied,thus providing molecular information for further studies on the effect of Ct CHS on flavonoids production.