Adoyo P A, Lea I A, Richardson R T, Widgren E E, O'Rand M G
Department of Cell Biology and Anatomy, University of North Carolina, Chapel Hill 27599, USA.
Mol Reprod Dev. 1997 May;47(1):66-71. doi: 10.1002/(SICI)1098-2795(199705)47:1<66::AID-MRD9>3.0.CO;2-O.
In this study, cDNAs encoding the sperm protein Sp17 from the baboon (Papio papio) have been cloned and sequenced. Three clones, differing in the lengths of their 3' untranslated regions, were identified, which were encoded by mRNA transcripts of 0.8-1.35 kb. The open reading frame encodes 163 amino acids with a predicted molecular mass of 18.8 kDa. The baboon Sp17 protein sequence is 97% identical to human Sp17 but differs significantly by the addition of 12 amino acids at the C-terminal, providing an additional potential protein kinase C phosphorylation site. Northern blot analysis demonstrated that the baboon Sp17 mRNA was specific to the baboon testes and was not detected in the ovary, placenta, or any of the other somatic tissues tested. Western blot analysis using anti-Sp17 antibodies demonstrated that the native baboon sperm Sp17 protein consists of a doublet with an apparent M(r) of 26.5 and 27.2 kDa. Immunocytochemical staining of baboon testis with anti-Sp17 antibodies demonstrated Sp17 in spermatocytes, spermatids, and spermatozoa within the seminiferous epithelium. No specific staining was observed on spermatogonia, Sertoli cells, Leydig cells, or other somatic cell types.
在本研究中,已克隆并测序了编码狒狒(巴氏狒狒)精子蛋白Sp17的cDNA。鉴定出三个3'非翻译区长度不同的克隆,它们由0.8 - 1.35 kb的mRNA转录本编码。开放阅读框编码163个氨基酸,预测分子量为18.8 kDa。狒狒Sp17蛋白序列与人类Sp17的序列一致性为97%,但在C末端额外添加了12个氨基酸,这提供了一个额外的潜在蛋白激酶C磷酸化位点。Northern印迹分析表明,狒狒Sp17 mRNA在狒狒睾丸中具有特异性,在卵巢、胎盘或任何其他测试的体细胞组织中均未检测到。使用抗Sp17抗体的Western印迹分析表明,天然狒狒精子Sp17蛋白由一条双峰带组成,表观分子量为26.5和27.2 kDa。用抗Sp17抗体对狒狒睾丸进行免疫细胞化学染色,结果显示生精上皮中的精母细胞、精子细胞和精子中有Sp17。在精原细胞、支持细胞、间质细胞或其他体细胞类型中未观察到特异性染色。