中文摘要：

目的：构建小鼠Spag6L基因真核表达质粒,并研究其在细胞内的表达与定位。方法：以小鼠睾丸cDNA基因文库为模版,PCR扩增Spag6L基因全长序列,测序正确后亚克隆至pEGFP-N2和pcDNA3真核表达载体中并酶切鉴定,将构建成功的重组表达质粒转染至COS-7与CHO细胞中,Western blot法检测COS-7细胞中SPAG6L蛋白表达,激光共聚焦扫描显微镜观察SPAG6L蛋白在CHO细胞内的定位。结果：重组质粒pEGFPSpag6L和pcDNA3-Spag6L经双酶切后产生的1.5kb的目的插入片段,经测序证实与Spag6L基因一致。GFPSPAG6L融合蛋白分子质量为82kU,SPAG6L蛋白分子质量为56kU。SPAG6L蛋白在CHO细胞中主要定位于细胞质,以微管和囊泡表达为主。结论：构建的pEGFP-Spag6L和pcDNA3-Spag6L真核表达质粒成功,为进一步探究Spag6L基因与Spag6基因的关系以及Spag6L基因在精子发生中的功能与作用奠定了基础。

英文摘要：

Objective： To construct eukaryotic plasmids expressing mouse SPAG6L protein, and investi gate its expression and localization in transfected mammaliam coding sequence was amplified by ploymerase chain reaction （ FP-N2 and pcDNA3 vector, respectively. After sequencing, t fected into CHO and COS-7 cells, and expression and cellu cells. Methods. The mouse Spag6L PCR） and subcloned into the pEG- he constructed plasmids were trans- lar localization of SPAG6L protein were determined by Western blot and immunofluorescence staining. Results. Mouse Spag6L was successfully cloned into the pEGFP-N2 and pcDNA3 vectors. Expression of SPAG6L and GFP- tagged SPAG6L fusion protein was confirmed, with the relative molecular weight of 56 kU and 82 kU respectively. SPAG6L protein mainly is localized in the cytoplasmic vesicles and microtu bules. Conclusion： The successful construction of the plasmids expressing SPAG6L lays a founda tion to further study its roles in vitro.