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林业科学 ›› 2007, Vol. 43 ›› Issue (02): 64-67.doi: 10.11707/j.1001-7488.20070211

• 论文及研究报告 • 上一篇    下一篇

松材线虫纤维素酶的免疫学分析

张奇1 李海燕2 白钢1 熊惠龙2 杨文博1   

  1. 1.南开大学生命科学学院,天津300071;2.国家林业局森林病虫害防治总站,沈阳110034
  • 收稿日期:2005-07-28 修回日期:1900-01-01 出版日期:2007-02-25 发布日期:2007-02-25

Immunological Analysis of Bursaphelenchus xylophilus Pathogenous Cellulase

Zhang Qi1,Li Haiyan2,Bai Gang1,Xiong Huilong2,Yang Wenbo1   

  1. 1. College of Life Science, Nankai University Tianjin 300071; 2. General Station of Forest Pest Control, State Administration of Forestry Shenyang 110034
  • Received:2005-07-28 Revised:1900-01-01 Online:2007-02-25 Published:2007-02-25

摘要:

制备松材线虫特定纤维素酶的多克隆抗体(Anti-BXC60),通过SDS-PAGE、纤维素酶酶染分析、免疫印迹以及ELISA方法考察该抗血清的特异性。进一步利用抗松材线虫全蛋白抗体(Anti-BX)和Anti-BXC60抗体,建立线虫分泌性蛋白在PVDF膜上双重染色的方法。双重染色分析的结果表明:松材线虫主要通过分泌孔向体外分泌大量的分泌蛋白,而纤维素酶的分泌方式与其他植物线虫相似,是通过口针向外分泌。

关键词: 松材线虫, 纤维素酶, 双重染色

Abstract:

After producing mouse anti-cellulase BXC60 polyclonal antibody (anti-BXC60), the specificity of Anti-BXC60 was verified by SDS-PAGE, cellulase staining, western blot and ELISA method. Further, we established a double staining method with rabbit anti-Bursaphelenchus xylophilus serum (Anti-BX) and Anti-BXC60, which could be used for analyzing secretory proteins on PVDF membrane. The result of double staining assay showed that the pine wood nematodes secreted some solute proteins through secretion pore, and cellulase was secreted from stylet that was the same as other plant nematodes.

Key words: Bursaphelenchus xylophilus, cellulase, double staining