Zhang Xiao-dong, Gou Da-wei, Miao Shi-ying, Zhang Jian-chao, Zong Shu-dong, Wang Lin-fang
State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2003 Jun;25(3):289-93.
To isolate and identify the differentially expressed genes in spermatogenesis for the understanding molecular mechanism of spermatogenesis.
Screening of the cDNA library, Northern blot, expression and purification in E. coli with GST expression system, immunocytochemical staining of testis sections were used.
(1) A cDNA fragment designated as RSD-7 was isolated from rat testis cDNA library. It was 1,238 bp in length, coding a protein of 232 amino acids with the GenBank accession number AF315467. The encoding protein of RSD-7 cDNA had a Ubiquitin-like domain. (2) Northern blot indicated that RSD-7 was uniquely expressed in rat testis, and in the testis RSD-7 emerged on the 30th postnatal day and expressed until 120th postnatal day. (3) Expression and purification of RSD-7 protein in E. coli with GST expression system and were used to obtain anti-RSD-7 antibody. (4) Immunolocalization of RSD-7 in rat testis revealed that it is expressed only in Sertoli cells.
Transcription pattern of RSD-7 and localization of RSD-7 protein in testis have been made, which established the base for the functional study of RSD-7.
分离并鉴定精子发生过程中差异表达的基因，以了解精子发生的分子机制。
采用筛选cDNA文库、Northern印迹、利用GST表达系统在大肠杆菌中表达和纯化以及睾丸切片免疫细胞化学染色等方法。
（1）从大鼠睾丸cDNA文库中分离出一个命名为RSD - 7的cDNA片段。其长度为1238 bp，编码一个含232个氨基酸的蛋白质，GenBank登录号为AF315467。RSD - 7 cDNA的编码蛋白具有一个泛素样结构域。（2）Northern印迹表明RSD - 7仅在大鼠睾丸中特异性表达，且在睾丸中RSD - 7于出生后第30天出现并持续表达至出生后第120天。（3）利用GST表达系统在大肠杆菌中表达并纯化RSD - 7蛋白，以此获得抗RSD - 7抗体。（4）RSD - 7在大鼠睾丸中的免疫定位显示它仅在支持细胞中表达。
已明确RSD - 7的转录模式及其在睾丸中的蛋白定位，为RSD - 7的功能研究奠定了基础。
