热激条件下家蚕血液30K蛋白磷酸化研究

Study on 30K Protein Phosphorylation of Silkworm Haemolymph under Heat Shock

本试验旨在研究热激条件下家蚕血液磷酸化蛋白的表达变化,为深入研究其功能提供参考。试验设对照组和热激组,每组选取五龄第3天家蚕幼虫各30头(雌雄各占1/2),在40℃条件下分别热激处理0、10 min,然后利用双向电泳、Pro-Q Diamond染色、质谱技术对表达差异的磷酸化蛋白进行研究。结果表明:1)银染图谱30 ku区积累了大量蛋白,经Pro-Q Diamond染色后发现它们属于磷酸化蛋白。2)经过多次取样和质谱鉴定,最终获得了3个30K蛋白,分别是PBMHPC-6、PBMHPC-19和PBMHPC-12。3)热激处理10 min后,3个30K蛋白的表达量与热激处理0 min相比显著上调(P<0.05)。由此得出,五龄家蚕血液富含30K蛋白,它们不仅在脂质运输方面承担着重要的角色,而且很可能参与热激应答。

This experiment was conducted to investigate the expression changes of phosphorylated protein in silkworm haemolymph under heat shock,and provided a reference for further study of its function. Two groups were designed,and 30 fifth-instar silkworms（ half male and half female） were randomly divided in each group,and treated with heat shock under 40 ℃ for 0（ control group） and 10 min（ heat shock group）,respectively. The differentially expressed phosphorylation proteins were studied utilizing two-dimensional electrophoresis,Pro-Q Diamond dye and mass spectrometry. The results showed as follow s： 1） amount of proteins were accumulated in the 30 ku area of silver staining map and revealed as phosphorylated proteins by Pro-Q Diamond dyeing. 2） Three 30 K proteins were identified as PBMHPC-6,PBMHPC-19 and PBMHPC-12 by multiple sampling and mass spectrometry identification. 3） Compared with 0 min heat shock,the expression levels of three 30 K proteins were significantly up-regulated after 10 min heat shock（ P〈 0.05）. It is concluded that fifthinstar silkworm haemolymph are rich in 30 K proteins. These proteins not only play important roles in lipid transport,but also may involve in heat shock response.