摘要
目的研究HSD13蛋白的组织表达定位及其对细胞周期的影响。方法利用王琳芳教授实验室构建的人睾丸特异ESTs文库,通过电子克隆方法获得一新基因命名为HSD13。通过Northern和Western印迹方法检测HSD13蛋白在不同组织中的表达,并用免疫组化观察其在小鼠睾丸组织中的表达定位。通过构建HSD13过表达的CHO稳定细胞系,用双胸苷阻断法并结合流式细胞术检测对细胞周期的影响。结果电子克隆获得HSD13基因。HSD13蛋白在睾丸组织中高表达,主要定位于生精细胞中的精原细胞和初级精母细胞。成功构建HSD13过表达和对照的CHO稳定细胞系,过表达HSD13蛋白可以显著加快细胞G2/M期进程。结论 HSD13蛋白于睾丸中精子发生早期表达,其过表达能够加速细胞周期进程。
Objective The localization of HSD13 in tissues and its effect on cell cycle were investigated. Methods HSD13 was collected by electronic cloning method. The expression of HSD13 in different tissues was detected by Northern blot and Western blot. The localization of HSD13 in testis was identified by immunohistochemistry. Doub- le thymidine block method in combination with flow cytometry was used to study the effect of HSD13 overexpression on cell cycle. Results The sequence of HSD13 was obtained by electronic cloning method. HSD13 was highly ex- pressed control process and its in testis, and was mainly located in spermatogonia and primary spermatocytes. HSD13 overexpression and CHO stable cell lines were constructed. Overexpression of HSD13 significantly accelerated G2/M phase in cell cycle. Conclusion HSD13 is highly expressed in spermatogonia and primary spermatocytes of testis, overexpression may accelerate cell cycle progression.
出处
《基础医学与临床》
CSCD
北大核心
2013年第2期179-183,共5页
Basic and Clinical Medicine
基金
国家重点基础研究发展项目(973计划)(2011CB944302)
国家重点实验室专项基金(2060204)
