Wang L F, Zhu H D, Miao S Y, Cao D F, Wu Y W, Zong S D, Koide S S
National Laboratory of Medical Molecular Biology, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, PRC.
Arch Androl. 1999 Mar-Apr;42(2):71-84. doi: 10.1080/014850199262904.
A 20-kDa sperm membrane protein cDNA, designated as RSD-1, was isolated by epitope selection from a rat testis lambda gtll expression library. RSD-1 was used as a probe to screen a human testis lambda ZAPII cDNA expression library. A cDNA designated as BS-63 was isolated and found to consist of 1933 bp with an open reading frame of 1824 bp and assigned the accession number U64675 by GenBank. The deduced polypeptide consisted of 608 amino acid residues containing XFXFG or FG motifs that are characteristic of nuclear pore complex (NPC) proteins and act as potential binding sites for Ran. The N-terminal region has high homology with RanBP2/Nup358, a nucleoporin component, showing that BS-63 is a member of the NPC family. Northern blot analysis of mRNAs prepared from various human tissues shows that BS-63 is transcribed in two forms: 6.0 and 8.5 kb. The 8.5-kb transcript was present in low amounts in several somatic tissues, whereas the 6.0-kb transcript is expressed only in testis. In situ hybridization analysis of human testis sections showed that BS-63 mRNA is expressed only in germ cells at all stages of spermatogenesis. Sertoli cells did not transcribe the gene.
通过表位选择从大鼠睾丸λgtll表达文库中分离出一个20 kDa的精子膜蛋白cDNA,命名为RSD - 1。用RSD - 1作为探针筛选人睾丸λZAPII cDNA表达文库。分离出一个命名为BS - 63的cDNA,发现其由1933 bp组成,开放阅读框为1824 bp,GenBank登录号为U64675。推导的多肽由608个氨基酸残基组成,含有XFXFG或FG基序,这些基序是核孔复合体(NPC)蛋白的特征,可作为Ran的潜在结合位点。N端区域与核孔蛋白成分RanBP2/Nup358具有高度同源性,表明BS - 63是NPC家族的成员。对从各种人体组织制备的mRNA进行Northern印迹分析表明,BS - 63以两种形式转录:6.0 kb和8.5 kb。8.5 kb的转录本在几种体细胞组织中含量较低,而6.0 kb的转录本仅在睾丸中表达。人睾丸切片的原位杂交分析表明,BS - 63 mRNA仅在精子发生各阶段的生殖细胞中表达。支持细胞不转录该基因。