特种经济动物营养 Special Economic Animal Nutrition

热激条件下家蚕血液30K蛋白磷酸化研究

  • 李娜 ,
  • 贾漫丽 ,
  • 杨贵明 ,
  • 李季生
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  • 1. 承德医学院蚕业研究所, 河北省高校特产蚕桑应用技术研发中心, 承德 067000;
    2. 浙江大学动物科学学院, 杭州 310029

收稿日期: 2015-12-18

  网络出版日期: 2016-06-27

基金资助

河北省教育厅资助项目(Q2012026,ZD2016044)

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

  • LI Na ,
  • JIA Manli ,
  • YANG Guiming ,
  • LI Jisheng
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  • 1. Sericulture Institute of Chengde Medical University, Hebei Universities R & D Centre for Sericulture and Specialty Enabling Technologies, Chengde 067000, China;
    2. College of Animal Sciences, Zhejiang University, Hangzhou 310029, China

Received date: 2015-12-18

  Online published: 2016-06-27

摘要

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。

本文引用格式

李娜 , 贾漫丽 , 杨贵明 , 李季生 . 热激条件下家蚕血液30K蛋白磷酸化研究[J]. 动物营养学报, 2016 , 28(6) : 1720 -1725 . DOI: 10.3969/j.issn.1006-267x.2016.06.012

Abstract

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 fifth-instar 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.

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