家蚕雌性附腺及其Ng突变体的蛋白质组差异研究

家蚕雌蛾性附腺在化蛾前2到3天开始大量分泌胶状粘性蛋白,其贮存部迅速地膨大,而其Ng突变体的雌蛾性附腺不能正常分泌胶状粘性物质.分别对家蚕(Bombyx mori)的正常及Ng突变体雌蛾性附腺分泌部组织的蛋白质进行提取,并采用双向凝胶电泳和计算机辅助分析方法,对提取的蛋白质混合物进行分离和比较分析,并对主要差异表达的蛋白质用质谱鉴定.实验结果表明,用银染法,平均每张电泳图谱可以分离约700个蛋白质点,其中大部分的蛋白质点分布在pH 4～8范围内,其分子质量主要集中在30～70 ku区域.比较分析发现一些差异表达蛋白,其中No2,3蛋白质点经质谱鉴定为肌动蛋白A3,该蛋白质只在化蛹后期正常雌性附腺组织中特异表达,而Ng突变体中肌动蛋白A3的缺失,暗示了肌动蛋白A3可能与家蚕雌性附腺的胶状粘性物质的胞外分泌有关.

Studies on The Proteome of Colleterial Gland and Its Ng Mutant of Silkworm (Bombyx mori )Using Two-dimensional Electrophoresis and Mass Spectrometry

The colleterial gland in the silkworm (Bombyx mori) grew gradually until 2 days before emergence, and markedly enlarged due to the accumulation of glue like substances (mainly including 85% water and 11% proteins).However, the Ng mutant female moth only secreted a small amount of glue-like substance and laid loose eggs naturally.High-resolution two-dimensional polyacrylamide gel electrophoresis, followed by computer-assisted analysis was used to screen the proteins pattern between normal and Ng mutant colleterial gland to find quantitative and qualitative difference in proteins expression. Protein spots were resolved in the secretory region of colleterial gland of silkworm and more than 700protein spots were resolved and most of the proteins were distributed in the area from 30 ku to 70 ku and pH 4 ～ 8. Through the comparison and analysis, it was found that 3 proteins were only expressed in the later pupae stage and moth stage.However, these proeins have no expression in the Ng mutant especially spot No. 2 and 3. These differentially expressed proteins were actin identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. The results indicated that the actins participate or regulate the exocytosis of colleterial gland and other differential expressed proteins might be having some relations with the glue-like proteins secretion.