Ma Quanhong, Wang Haikun, Guo Rui, Wang Huizhen, Ge Yehua, Ma Jing, Xue Shepu, Han Daishu
Department of Cell Biology, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing, China.
Mol Reprod Dev. 2006 Sep;73(9):1075-83. doi: 10.1002/mrd.20485.
Full-length cDNA of a novel mouse gene upregulated in late stages of spermatogenic cells was cloned from mouse testis using overlapping RT-PCR and RACE. The mRNA of the gene was expressed mainly in diplotene/pachytene spermatocytes, round and elongating spermatids. We named this gene as SRG-L (Spermatogenesis Related Gene expressed in late stages of spermatogenic cells, GenBank Accession No. AY352586). The tissue-specific analysis showed a higher expression level in testis and spleen. The gene is mapped on chromosome 8q33.1 and contains 18 exons. The full-length of cDNA is 2,843 bp with an open reading frame (ORF) of 2,625 bp that encodes a 104 kDa protein (874 amino acids) with a putative transmembrane region. The bioinformatics analysis revealed that the SRG-L has two conserved regions, transglutaminase-like homologues domain and D-serine dehydratase domain, rich phosphorylation sites and methylation sites. The SRG-L protein was detected in diplotene/pachytene spermatocytes and spermatids by immunohistochemical staining and Western blot. The results suggest that SRG-L may play definite roles regulating differentiation of germ cells during spermatogenesis, particularly during meiosis and spermiogenesis.
利用重叠RT-PCR和RACE技术从小鼠睾丸中克隆出一个在生精细胞后期上调的新型小鼠基因的全长cDNA。该基因的mRNA主要在双线期/粗线期精母细胞、圆形和伸长型精子细胞中表达。我们将该基因命名为SRG-L(在生精细胞后期表达的生精相关基因,GenBank登录号为AY352586)。组织特异性分析显示,该基因在睾丸和脾脏中的表达水平较高。该基因定位于8号染色体q33.1,包含18个外显子。cDNA全长2843bp,开放阅读框(ORF)为2625bp,编码一个104kDa的蛋白质(874个氨基酸),具有一个推定的跨膜区域。生物信息学分析表明,SRG-L有两个保守区域,转谷氨酰胺酶样同源结构域和D-丝氨酸脱水酶结构域,富含磷酸化位点和甲基化位点。通过免疫组织化学染色和蛋白质免疫印迹法在双线期/粗线期精母细胞和精子细胞中检测到了SRG-L蛋白。结果表明,SRG-L在精子发生过程中,特别是在减数分裂和精子形成过程中,可能在调节生殖细胞分化方面发挥一定作用。