银鲫精巢特异的Ly-6/uPAR相关蛋白的分子克隆及其特征分析  

MOLECULAR CLONING AND EXPRESSION ANALYSIS OF TESTIS-SPECIFIC Ly-6/u PAR RELATED PROTEIN IN GIBEL CARP

作　　者：王伟[1,2] 刘珍 周莉[1] 李志[1] 桂建芳[1] 

机构地区：[1]中国科学院水生生物研究所淡水生态与生物技术国家重点实验室,武汉430072 [2]中国科学院大学,北京100049

出　　处：《水生生物学报》2015年第3期441-448,共8页Acta Hydrobiologica Sinica

基　　金：国家自然科学基金(31123001);淡水生态与生物技术国家重点实验室(2011FBZ17)资助

摘　　要：Ly-6/u PAR基因超家族(Ly-6 SF)成员广泛地存在于后生动物中,开展该家族相关功能基因研究具有重要的意义。研究从银鲫(Carassius auratus gibelio)中鉴定到一个该家族新成员,c DNA全长为570 bp,其中开放阅读框长度为300 bp,编码99个氨基酸,生物软件预测该蛋白含有一个LU结构域,不含GPI锚信号序列,N端含有信号肽,表明其可能为Ly-6基因超家族中分泌型蛋白。组织表达分析显示,该基因只在银鲫精巢中特异表达,且又是Ly-6基因超家族中一员,因此将其命名为银鲫精巢特异的Ly-6/u PAR相关蛋白(Carassius auratus gibelio testis-specific Ly-6/u PAR related protein,简称Cag Tslurp)。原位杂交结果显示,该基因在银鲫精巢的精原细胞,初级精母细胞以及次级精母细胞中表达,精子细胞中存在少量的表达,而在体细胞中不表达。这种精巢特异的表达模式,暗示Cag Tslurp在银鲫精子发生中可能发挥了作用。The members of Ly-6/u PAR superfamily distribute widely in many metazoans. All proteins from this family have at least one conserved Ly-6/u PAR（LU） domain which is composed of about 70 to 100 amino acids, and contain 8 to 10 cysteine residues with a defined disulfide-bonding pattern. The members of Ly-6 family are classified into two subfamilies, namely GPI-anchored membrane proteins and secreted proteins, according to whether they possess the glycophosphatidyl inositol（GPI）-anchored signal sequence in the C-terminus. In this study, we cloned and characterized a new member of Ly-6 superfamily in gibel carp（Carassius auratus gibelio）. The full-length sequence of this gene consisted of 570 base pairs that encoded 99 amino acids and had only one LU domain. Our analysis using on-line bioinformatic software showed that this protein might have one signal peptide but no GPI-anchor signal sequence, thus could be a type of secreted protein. We conducted RT-PCR to detect its expression in different tissues and found that this protein was specifically expressed in testis. According to its characteristics, we named the gene Carassius auratus gibelio testisspecific Ly-6/u PAR related protein（Cag Tslurp）. In situ hybridization revealed the expression of Cag Tslurp in spermatogonia, primary spermatocytes, secondary spermatocytes and spermatids, but not in somatic cells. These results indicated that the Cag Tslurp gene might play a role in spermatogenesis of gibel carp.

关 键 词：银鲫 Ly-6基因超家族 CagTslurp 精巢特异 精子发生 原位杂交 

分 类 号：Q344.1[生物学—遗传学]