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Molecular cloning and characterization of SRG-L, a novel mouse gene developmentally expressed in spermatogenic cells

机译:SRG-L的分子克隆和鉴定,一种在生精细胞中发育表达的新型小鼠基因

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摘要

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.
机译:使用重叠RT-PCR和RACE技术从小鼠睾丸中克隆了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主要表达于二倍体/粗种精细胞,圆形和伸长的精细胞中。我们将该基因命名为SRG-L(在生精细胞的晚期表达的生精相关基因,GenBank登录号AY352586)。组织特异性分析显示睾丸和脾脏中的表达水平较高。该基因位于8q33.1染色体上,包含18个外显子。 cDNA的全长为2,843 bp,开放阅读框(ORF)为2,625 bp,编码一个104 kDa的蛋白(874个氨基酸),并带有一个跨膜区。生物信息学分析表明,SRG-L具有两个保守区域,即转谷氨酰胺酶样同源结构域和D-丝氨酸脱水酶结构域,具有丰富的磷酸化位点和甲基化位点。通过免疫组织化学染色和Western印迹检测到二倍体/粗种精细胞和精细胞中的SRG-L蛋白。结果表明SRG-L可能在精子发生过程中,特别是在减数分裂和精子发生过程中,起着调节生殖细胞分化的作用。

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