中心蛋白3定位于游仆虫细胞中形成棘毛的基体上

中心蛋白与中心体(或基体)有着密切关系,作为一种细胞骨架蛋白,形成原生动物巨大的细胞骨架网络;为了研究从八肋游仆虫细胞中克隆到的一种中心蛋白基因(EoCen3)的功能,我们构建了八肋游仆虫细胞大核人工染色体pBTub-Tel,定位分析该基因所表达的中心蛋白在细胞中的分布和行为.该人工染色体利用密码子优化的增强型绿色荧光蛋白基因作为标记,在八肋游仆虫细胞中实现高表达,避免用抗生素作为标记对该细胞的喂养产生影响.定位结果显示,该类中心蛋白特异定位于游仆虫形态发生过程中的基体上.在八肋游仆虫无性生殖细胞分裂的起始阶段基体开始发育,首先形成第一个基体,Eo-Cen3定位于新形成的基体中;随后基体开始复制,EoCen3定位于正在复制的基体中.在细胞发育过程中绿色荧光标记的基体成倍增加,说明EoCen3与参与棘毛形成的基体的复制有一定的关系.

Centrin 3 is Localized Restrictedly in Duplicating Basal Body Responsible for Formation of Cirri in Euplotes

Centrins are generally associated with the centrosome/basal body, and, as ubiquitous cytoskeletal related proteins,form large cytoskeletal networks in protist. To make sure the function of a centrin gene (EoCen3) that was cloned from Euplotes octocarinatus ,its distribution and behavior in one ciliate species, E. octocarinatus, using a novel expression veetor,pBTub-Tel,which were developed specifically for ciliates Euplotes cell were studied. Codon-opti-mized EGFP(EGFP-Eo)was used as marker, and avoiding using antibiotics which may affect the cell culture by feed-ing. The localization results indicated that EoCen3 was detected specifically within the basal bodies during morpho-genesis of Euplotes. While the basal body generated de novo at the initiate stage of division, EoCen3 occured within duplicating basal bodies. Double increase of basal body marked with green fluorescence during the morphogenesis suggests that EoCen3 may be related to the duplication of basal body for formation of cirri.