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目的通过筛选人睾丸特异基因及对其结构和功能的研究，为阐明精子发生与成熟提供新的线索。方法采用筛选人睾丸表达文库、快速扩增ｃＤＮＡ５′末端、Ｎｏｒｔｈｅｒｎ印迹分析、荧光原位杂交（ＦＩＳＨ）以及基因重组等技术。结果（１）获得一新的长度为２２０９ｂｐ的ｃＤＮＡ，编码７００个氨基酸，将该基因命名为ＢＳ－６３。Ｇｅｎ－Ｂａｎｋ注册号为Ｕ６４６７５。（２）ＢＳ－６３Ｎ端具有核孔蛋白的特征性结构元件“ＸＦＸＦＧ”或“ＦＧ”以及与Ｒａｓ相关核蛋白（Ｒａｎ）结合的典型特征。（３）Ｎｏｒｔｈｅｒｎ印迹分析显示，ＢＳ－６３基因有６．０和８．５ｋｂ两种转录形式，其中６．０ｋｂ为睾丸组织特异性转录本。（４）ＦＩＳＨ显示，ＢＳ－６３基因定位于２ｑ１１．２～１２。（５）ＢＳ－６３ｃＤＮＡ在大肠杆菌中获得高效表达，ＳＤＳ－ＰＡＧＥ显示，表达蛋白的相对分子质量约为８００００。（６）体外功能实验表明，该表达蛋白可被蛋白激酶Ｃ及细胞周期依赖性蛋白激酶Ⅱ磷酸化。结论ＢＳ－６３可能是一种新的睾丸特异核孔蛋白。
Objective By screening human testis-specific genes and their structure and function, to provide new clues to elucidate the spermatogenesis and maturation. Methods Screening human testis expression library, rapid amplification of cDNA 5 ’end, Northern blot analysis, fluorescence in situ hybridization (FISH) and gene recombination techniques. Results (1) A new 2209bp cDNA was obtained, encoding 700 amino acids. The gene was named BS-63. Gen-Bank registration number is U64675. (2) BS-63N possesses the characteristic features of nuclear pore protein “XFXFG” or “FG” as well as the typical features associated with Ras-associated nucleoprotein (Ran). (3) Northern blot analysis showed that the BS-63 gene has two transcriptional forms of 6.0 and 8.5kb, of which 6.0kb is testicular tissue-specific transcript. (4) FISH shows that BS-63 gene is located in 2q11.2 ~ 12. (5) BS-63cDNA was highly expressed in E.coli. SDS-PAGE showed that the relative molecular mass of the expressed protein was about 80,000. (6) In vitro functional experiments show that the expressed protein can be phosphorylated by protein kinase C and cell cycle dependent protein kinase Ⅱ. Conclusion BS-63 may be a new testicular-specific nuclear porin.