家蚕孤雌生殖相关蛋白PGL的鉴定与生物信息学分析

应用二维凝胶电泳(2DE)技术分离有性生殖蚕卵与孤雌生殖蚕卵的差异蛋白,通过基质辅助激光解吸电离串联飞行时间质谱(MALDI-TOF-TOFMS)分析获得大量小肽的序列特征。与已构建的家蚕cDNA文库及蛋白质数据库进行比对,确定一个在家蚕孤雌生殖过程中表达量显著上调的差异蛋白为磷脂酰肌醇蛋白聚糖(phosphatidylinositol glycan,PGL)。为了探讨PGL蛋白在家蚕孤雌生殖发生中的作用,采用cDNA末端快速扩增(RACE)技术扩增目的基因5′端片段,获得PGL蛋白基因的全长cDNA,生物信息学分析显示该基因cDNA全长1038bp,编码345个氨基酸,其编码蛋白理论分子质量为39.516kD,等电点为5.845,包含1个保守的GPI8结构域,信号肽概率为0.997,可能为分泌型蛋白,预测蛋白功能域包含酪蛋白激酶Ⅱ磷酸化、N糖基化、蛋白激酶C磷酸化等多个功能位点。依据PGL蛋白的理化性质、结构功能推测其有可能在不同程度上通过细胞信号转导途径参与了家蚕孤雌生殖发生过程并发挥重要作用。

Identification and Bioinformatic Analysis of a Parthenogenesis Associated Protein Phosphatidylinositol Glycan in Bombyx mori

Two-dimension gel electrophoresis (2DE) technique was used to identify differential protein spots between Bombyx mori eggs from normal sexual reproduction and those from parthenogenetic reproduction.After these protein spots were analyzed by MALDI-TOF-TOF MS,a large number of short peptide sequences were obtained.Alignments with sequences in a constructed cDNA library and in predicted protein database of Bombyx mori led to the identification of a phosphatidylinositol glycan protein (PGL) of which expression...