银鲫卵母细胞特异表达凝集素的抗体制备及其定位分析

POLYCLONAL ANTIBODY PREPARATION AND IMMUNOFLUORESENCE LOCALIZATION OF GIBEL CARP OOCYTE2SPECIFIC LECTIN

  • 摘要: 银鲫卵母细胞特异的凝集素(Gibel carp oocyte-specific lectin,GOL)是经差减杂交筛选出来的。本研究采用原核表达方法,利用Pinpoint-T载体表达了该凝集素的部分肽段(从22到214个氨基酸),并经纯化用来免疫家兔制备多克隆抗体。采用免疫荧光共定位方法,证明该凝集素是卵母细胞中皮质颗粒的成分之一。

     

    Abstract: Lectins were defined as proteins that recognize specific carbohydrate structures and agglutinate cells by binding to cell surface glycoproteins and glycoconjugates. Many kinds of different lectins have been purified from fish eggs.Lectins may have a variety of biological function in fish eggs,such as block of polyspermy,regulation of carbohydrates metabolism,participation in the formation of fertilization envelope after binding with glycoproteins,antibacterial effect and opsonization of pathogens. Therefore, lectins are important components in oocytes and play important roles. Cortical granules were as a kind of secretory vesicles that unique to oocyte. They are synthesized and accumulated during oogenesis,translocated to the cell surface before fertilization and exocytosed after fertilization. Cortical granule contents have diverse components,such as proteases and lectins,and play important roles in egg fertilization and early embryogenesis.

     

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