CAJ | 학술논문

衣藻RS P3是一个鞭毛结构蛋白，目前有关RS P3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RS P3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RS P3与微管、内质网以及高尔基体的关系。结果表明，小鼠RS P3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C H O细胞中表达的小鼠RS P3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。表明RS P3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。
의조RS P3시일개편모결구단백，목전유관RS P3상관연구주요집중재의조상，이대기재포유동물중적동원기인적단백정위급공능연구심소。본연구대각개물충적RS P3안기산서렬진행비대분석，재용SMART 연건예측소서RSP3결구역적기출상구건료소서RSP3진핵표체재체，병재CHO세포중성공표체，응용형광공정위기술연구소서RS P3여미관、내질망이급고이기체적관계。결과표명，소서RS P3비의조RSP3재N‐단다출127개안기산，량자도함유일개RSP3결구역，RSP3결구역시일개미관결합결구역，이차재진화중고도보수。재C H O세포중표체적소서RS P3병불직접여미관결합，야불정위재내질망혹고이기체，진일보연구발현기취집분포우세포질기질중，이차주요분포재세포핵적주위。표명RS P3재진화중교위보수，단재포유동물중적분포화공능가능여재의조중유소불동。
Chlamydomonas reinhardtii RSP3 is the component of flagella .Previous studies mainly focus on Chlamydomonas flagella ,w hile the localization and function of the homologous gene on mammalians is not clear .We analysis the amnion acid sequence of RSP3 from different organism and predict the protein do‐main of RSP3 .Then we construct recombinant plasmid of the mouse RSP3 and expressed in the CHO cells and co‐labeled with microtubules ,endoplasmic reticulum or Golgi apparatus .The results show that the mouse RSP3 have another 127 amino acids in the N‐terminal compared to Chlamydomonas RSP3 .Both mouse and Chlamydomonas RSP3 have a RSP3 domain ,w hich regards as a conserved microtubule binding domain .Co‐labled analysis results suggest that the mammalian RSP3 does not bind to microtubules ,endo‐plasmic reticulum or Golgi apparatus ,it aggregates in the cytoplasm surround the nuclear .These indicated that the subcelluar localization and function of mammalian RSP 3 may be different from it in the Chlamydo‐monas .