小鼠RSP3基因序列分析及其亚细胞定位  

Sequence Analysis and Subcellular Localization of Mouse RSP3 Gene

作　　者：成欣然 颜润川 胡新德[1] 宋玲珍[1] 张伟[1] 张亚妹[1] 陈树林[1] 赵善廷[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《动物医学进展》2015年第2期1-7,共7页Progress In Veterinary Medicine

基　　金：西北农林科技大学人才引进专项基金(Z111021101)

摘　　要：衣藻RSP3是一个鞭毛结构蛋白,目前有关RSP3相关研究主要集中在衣藻上,而对其在哺乳动物中的同源基因的蛋白定位及功能研究甚少。本研究对各个物种的RSP3氨基酸序列进行比对分析,在用SMART软件预测小鼠RSP3结构域的基础上构建了小鼠RSP3真核表达载体,并在CHO细胞中成功表达,应用荧光共定位技术研究小鼠RSP3与微管、内质网及高尔基体的关系。结果表明,小鼠RSP3比衣藻RSP3在N端多出127个氨基酸,两者都含有一个RSP3结构域,RSP3结构域是一个微管结合结构域,而且在进化中高度保守。在CHO细胞中表达的小鼠RSP3并不直接与微管结合,也不定位在内质网或高尔基体,进一步研究发现其聚集分布于细胞质基质中,而且主要分布在细胞核的周围。说明RSP3在进化中较为保守,但在哺乳动物中的分布和功能可能与在衣藻中有所不同。Chlarnydornonas reinhardtii RSP3 is the component of flagella. Previous studies mainly focus on Ghlarnydornonas flagella, while the localization and function of the homologous gene on mammalians are not clear. We analyzed the amnion acid sequence of RSP3 from different organisms and predicted the protein domain of RSP3. Then we constructed recombinant plasmid of the mouse RSP3 and expressed in the CHO cells and co-labeled with microtuhules, endoplasmic reticulum or Golgi apparatus. The results showed that the mouse RSP3 has another 127 amino acids in the N-terminal compared to Chlarnydornonas RSP3. Both mouse and Chlamydornonas RSP3 have a RSP3 domain, which regards as a conserved microtubule binding domain. Co-labled analysis results suggested that the mammalian RSP3 does not bind to microtubules, endoplasmic reticulum or Golgi apparatus, it aggregates in the cytoplasm surround the nuclear. These indicated that the subcelluar localization and function of mammalian RSP3 may be different from it in the Chlarnydornonas.

关 键 词：RSP3 序列分析 亚细胞定位 内质网 高尔基体 

分 类 号：Q785[生物学—分子生物学]