摘要
目的 从家蝇蛹中分离纯化一种能结合半乳糖的凝集素 ,并研究其免疫和抑菌活性。方法 家蝇蛹凝集素提取纯化方法包括 :缓冲液浸提、戊二醛固定化红细胞吸附、亲和层析和凝胶过滤。以淋巴细胞转化、溶血空斑和抑菌试验研究此凝集素的性质。结果 本纯化方法使样品的血凝活力提高了 5 39倍 ,纯化后的家蝇蛹凝集素电泳测得其相对分子质量 (Mr)为 84× 10 3,由 3个亚单位组成 ,含有糖成分。纯化凝集素的浓度为 15 .6～ 10 0 0 μg ml时 ,具有刺激T、B细胞的作用 ,与对照组比较差异有显著性 (P <0 .0 1)。对大肠埃希菌 ,枯草芽胞杆菌和伤寒沙门菌的抑制活性分别为39 .8%、4 5 .3%和 6 6 .2 %。结论 家蝇蛹凝集素具有免疫调节功能和抑菌活性 ,为家蝇免疫机制的深入研究提供了资料。
Objective Isolation of a new galactose-specific lectin (MPL) from Musca domestic pupae and study of its immunoactivity and antibacterial activity. Methods The purification procedure entailed extraction with aqueous buffer, GRBC (glutaraldehydeized red blood cells) adsorption, affinity chromatography on Sepharose-4B and gel-filtration on SephadexG-200. The characteristic of MPL was demonstrated by lymphocyte transformation test, plaque-forming cell assay and antibacterial activity test. Results The purification method increased the lectin′s hemagglutination activity by 539 times. By SDS-PAGE, the molecular weight of MPL was 84kDa, it was made of three subgroups and contained carbohydrate structure. When the concentration of MPL was 15.6-1?000μg/ml, it stimulated T and B lymphocyte and had significant difference compared with the control ( P <0.01). Antibacterial activity of MPL against Escherichia coli, Bacillus subtilis and Salmonella typhi was 39.8%, 45.3% and 66.2%. Conclusion MPL is an antibacterial and immunoactive protein.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2003年第10期805-808,共4页
Chinese Journal of Microbiology and Immunology
基金
天津自然科学基金重点项目 (0 0 3 80 3 911)
