热激条件下家蚕血液30K蛋白磷酸化研究

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(P0.05)。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。

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

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 follows: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 30K 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 30K proteins were significantly up-regulated after 10 min heat shock (P＜0.05).It is concluded that fifthinstar silkworm haemolymph are rich in 30K proteins.These proteins not only play important roles in lipid transport,but also may involve in heat shock response.