水蕨中心体蛋白基因的克隆与原核表达

发布时间：2018-05-04 00:12

本文选题：中心体蛋白 + 克隆　；参考：《西北植物学报》2016年03期

【摘要】：蕨类植物的精子形成是一个运动细胞器重新发生的过程,但对其精子发生的分子机制仍不甚了解。中心体蛋白(centrin)是定位于细胞中心体上的一种高度保守的钙结合蛋白,被认为可能是最早参与运动细胞器发生的蛋白质。克隆鉴定centrin蛋白为进一步分析蕨类精子发生的分子机制奠定基础。该研究从蕨类模式植物水蕨(Ceratopteris thalictroides)中克隆到了水蕨centrin基因(CtCEN),其cDNA序列全长为1 077bp,结构分析表明CtCEN基因属于EF-hand超家族的centrin基因。Centrin的系统树分析发现,藻类、蕨类以及动物等具鞭毛游动精子的生物聚为一支,而被子植物聚为另一分支,表明centrin的功能可能与运动细胞器鞭毛发生有关。原核表达和Western blot分析表明,CtCEN基因表达的蛋白能与人源centrin抗体发生结合,这一结果证实克隆到了centrin基因。
[Abstract]:Spermatogenesis in ferns is a process of reoccurrence of motility organelles, but the molecular mechanism of spermatogenesis is still unknown. Centrosomal protein (CEP) is a highly conserved calcium-binding protein located on the centrosome of cells, which may be the earliest involved in the development of motor organelles. Cloning and identification of centrin protein lay a foundation for further analysis of the molecular mechanism of spermatogenesis in ferns. In this study, centrin gene CtCENN was cloned from the fern model plant Ceratopteris thalictroids. Its cDNA sequence was 1 077bp.The structural analysis showed that the CtCEN gene belongs to the EF-hand superfamily centrin gene .Centrin phylogenetic tree analysis found that algae. The spermatozoa with flagellum such as ferns and animals are clustered into one branch, while angiosperms form another branch, indicating that the function of centrin may be related to the development of the flagella of the motility organelles. Prokaryotic expression and Western blot analysis showed that the protein expressed by CtCEN gene could bind to human centrin antibody, which confirmed the cloning of centrin gene.
【作者单位】：上海师范大学生命与环境科学学院;上海资源植物功能基因组学重点实验室上海辰山植物园;
【基金】：上海市自然科学基金(13ZR1429700) 上海市科学技术委员会课题(14DZ2260400)
【分类号】：Q943.2

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