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草鱼血清中一种新的凝集素免疫因子
邵健忠, 项黎新
浙江大学生命科学学院 杭州310012
摘要:
于1998年5月–2000年3月,在浙江省杭州市采集草鱼,采用硫酸铵盐析和梯度聚丙烯酰胺凝胶电泳技术,从血清中分离出一种自然凝集素,对其理化和生物学性质进行研究。结果表明,该凝集素能够识别和凝集多种不同的红细胞、细菌和酵母,具有100000g离心不沉降,不被透析,耐热、耐酸、耐碱,抗DNase、RNase、SDS和乙醚,对胰蛋白酶、糖苷酶、糖氧化剂NaIO4和β-巯基乙醇敏感等特性,是一种分子量为67kD、等电点为6.2的糖蛋白分子。其活性能被半乳糖、乳糖、甘露糖和N-乙酰葡糖胺所抑制,其抗原特异性不同于草鱼凝集抗体IgM,表明两者是不同的物质。草鱼凝集素能显著地促进巨噬细胞的吞噬、杀菌能力,说明是一种重要的免疫活性因子。
关键词:  草鱼凝集素  分离纯化  理化性质  免疫活性
DOI:
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基金项目:国家自然科学基金资助项目39970589号;浙江省自然科学基金资助项目397300号
A NATURAL IMMUNOREACTIVE AGGLUTININ ISOLATED FROM SERUM OF GRASS CRAP (CTENOPHARYNGODON IDELLUS)
SHAO Jian-Zhong, XIANG Li-Xin
College of Life Science, Zhejiang University, Hangzhou, 310012
Abstract:
A naturally occurring immunoreactive agglutinin was isolated from serum of grass carp (Ctenopharyngodon idellus) by means of salting out in ammonium sulphate and gradient polyacrylamide gel electrophoresis were used during May, 1998 to March, 2000 in Hangzhou City, Zhejiang Province of China. Bio-physiochemical studies showed that the agglutinin has haem agglutinating activity for a variety of animal erythrocytes, bacteria and yeast cells. It could not be dialyzed and sedimented at 100,000g, exhibited stability to heat, pH=2, pH =10, DNase, RNase, SDS and lipid solvent aether, but sensitive to trypsin, glucosidase, sugar oxidant NaIO4 and β-mercaptoethanol. It was shown to be a glycoprotein with the molecular weight of 67kD and isoelectric point of 6.2. Its activity could be inhibited by galactose, lactose, mannose and N-acetylglucosamine respectively, and its antigenic characteristic was not the same as that of antibody (IgM) of grass crap, which indicated that this agglutinin was a substance different from antibody. That the phagocytic and bactericidal activity of grass carp macrophages could be significantly increased by this agglutinin, indicates that the agglutinin is probably an important immune factor in fish.
Key words:  Agglutinin of grass carp, Isolation, Bio- physiochemical characteristics, Immune activity
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