中文摘要：

目的：探讨精子相关抗原6（sperm associated antigen 6,Spag6）和T复合体相关睾丸表达3（T-complex associated testis expressed 3,TCTE3）之间的相互作用。方法：PCR法观察Tcte3在雄性小鼠各组织中的表达;以小鼠睾丸c DNA为模板,扩增Tcte3序列,构建Tcte3/p GADT7、Tcte3/p EGFPN2重组质粒;以Tcte3/p EGFPN2转染CHO细胞和COS-1细胞,免疫荧光染色及Western blot检测TCTE3在细胞中的定位和表达;以Spag6/p GBKT7和Tcte3/p GADT7进行酵母双杂交实验,提取酵母蛋白后采用Western blot检测TCTE3与SPAG6之间是否存在直接的相互作用。结果：Tcte3在睾丸、脑、肝、肾、肺、脾、肌肉中均有表达;酶切和测序鉴定证实Tcte3/p GADT7和Tcte3/p EGFPN2重组质粒均构建成功;体外实验发现TCTE3主要在CHO和COS-1细胞质中表达,当与SPAG6质粒共转染后,TCTE3被募集至细胞微管中表达;酵母双杂交实验显示TCTE3/SPAG6组合后,在缺少SD/-Leu/-Trp/-His的培养板上有菌落生成,Western blot显示菌落中TCTE3、SPAG6蛋白表达显著,分子量正确。结论：精子相关抗原SPAG6与TCTE3之间存在直接的相互作用,SPAG6可募集TCTE3至细胞微管中表达。

英文摘要：

Objective：To test the interactions between SPAG6 and Tcte3 proteins. Methods：The expression of Tcte3 was observed in different tissues of mice by PCR. The testis c DNA was used as the template to amplify Tcte3 coding sequence by PCR,following with the Tcte3/p GADT7 and Tcte3/p EGFPN2 recombinant plasmids＇ constructing. Immunofluorescent staining and Western blot were used to detect the location and protein express after transfecting Tcte3/p EGFPN2 into CHO cells and COS-1 cells. The interaction between SPAG6 and TCTE3 was detected by Western blot after yeast two hybrid for Tcte3/p GADT7 and SPAG6/PGBKT7. Results：Tcte3mainly expressed in testis,brain,liver,kidney,lung,spleen and muscle. The results of restriction enzyme digestion and sequencing showed the Tcte3/p GADT7 and Tcte3/p EGFPN2 recombinant plasmids were constructed successfully. TCTE3 mainly located in the whole cytoplasm in vitro,whereas it was recruited to microtubule after co-transfecting with SPAG6. In yeast two hybrid assay,the mating group of TCTE3/SPAG6 grow promoted colonies to grow in SD/-Leu/-Trp/-His plates,and Western blot assay had identified TCTE3 and SPAG6 proteins could express in these colonies. Conclusion：The directly interaction between SPAG6 and TCTE3 is confirmed,and SPAG6 could recruite TCTE3 to celluar microtubule.