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王伟, 刘珍, 周莉, 李志, 桂建芳. 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[J]. 水生生物学报, 2015, 39(3): 441-448. DOI: 10.7541/2015.59
引用本文: 王伟, 刘珍, 周莉, 李志, 桂建芳. 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[J]. 水生生物学报, 2015, 39(3): 441-448. DOI: 10.7541/2015.59
WANG Wei, LIU Zhen, ZHOU Li, LI Zhi, GUI Jian-Fang. MOLECULAR CLONING AND EXPRESSION ANALYSIS OF TESTIS-SPECIFIC Ly-6/uPAR RELATED PROTEIN IN GIBEL CARP[J]. ACTA HYDROBIOLOGICA SINICA, 2015, 39(3): 441-448. DOI: 10.7541/2015.59
Citation: WANG Wei, LIU Zhen, ZHOU Li, LI Zhi, GUI Jian-Fang. MOLECULAR CLONING AND EXPRESSION ANALYSIS OF TESTIS-SPECIFIC Ly-6/uPAR RELATED PROTEIN IN GIBEL CARP[J]. ACTA HYDROBIOLOGICA SINICA, 2015, 39(3): 441-448. DOI: 10.7541/2015.59

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/uPAR RELATED PROTEIN IN GIBEL CARP

  • 摘要: Ly-6/uPAR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, cDNA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/uPAR相关蛋白(Carassius auratus gibelio testis-specific Ly-6/uPAR related protein, 简称CagTslurp)。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CagTslurp在银鲫精子发生中可能发挥了作用。

     

    Abstract: The members of Ly-6/uPAR superfamily distribute widely in many metazoans. All proteins from this family have at least one conserved Ly-6/uPAR (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 testis-specific Ly-6/uPAR related protein (CagTslurp). In situ hybridization revealed the expression of CagTslurp in spermatogonia, primary spermatocytes, secondary spermatocytes and spermatids, but not in somatic cells. These results indicated that the CagTslurp gene might play a role in spermatogenesis of gibel carp.

     

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