草鱼干扰素的分离纯化及某些理化和生物学特性
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浙江大学生命科学学院

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国家自然科学基金资助项目( 草鱼非特异性免疫因子干扰素的研究和草鱼干扰素基因克隆及其在衣薄中表达的研究) , 39400100 和39970589 号。


Purification and some physico..chemical and biological characterization of Ctenocpharyngodon idellus interferon
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College of Life Science,Zhejiang University, Hangzhou 310012

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    摘要:

    通过DEAE..Sepharose 阴离子交换层析、S ephacryl S..200 凝胶层析、高效液相层析和梯度聚丙烯 酰胺凝胶电泳等技术, 对病毒诱生的草鱼血清干扰素进行了分离纯化。纯化的草鱼干扰素在SDS..聚丙烯酰 胺凝胶电泳中呈现单一组份, 分子量为38kD, 等电点为5. 25, 过碘酸..Schiff 试剂反应表现为典型的糖蛋白染 色特征。理化和生物学特性研究表明, 草鱼干扰素具有100 000g( 2h) 离心不沉降、耐热、在pH2- 10 范围内活 性稳定、抗核酸酶, 但对胰蛋白酶、糖苷酶和糖氧化剂NaIO4 敏感等性质, 在鱼类细胞中具有抑制不同病毒的 能力, 但在人、哺乳类、鸟类和贝类等细胞中无抗病毒作用, 表现出抗病毒的种属特异性, 与高等脊椎动物../ .. 干扰素的特性相一致。

    Abstract:

    Serum interferon of grass carp ( Ctenocpharyngodon idellus ) , which was induced by the haemorrhagic virus was purified by means of DEAE..Sepharose anion exchane chromatography, Sephacryl S..200 gel chromatography, high..performance liquid chromatography and gradient polyacrylamide gel electrophoresis. The purif ied grass carp interferon exhibited single component when analysed by the SDS polyacrylamid gel electrophoresis. It was a glycoprotein with the molecular weight of 38kD and the isoelectric point of 5. 25. Biochemical studies showed that the grass carp interferon was not sedimented at 100 000g for 2 hours, exhibited stability to heat of pH2~ 10, and resisting to DNase and RNase, but was sensitive to trypsin, Glucosidase and NaIO4. It was able to inhibit different kinds of virus in the cells of grass carp, but failed in human, mammalian, avian and shellf ish cells. The results showed that the grass carp interferon was specif ic to fish cells and viruses. Its characterization was consistant w ith that of high vertibrate ../ .. interferon.

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邵健忠.草鱼干扰素的分离纯化及某些理化和生物学特性[J].水产学报,2000,24(1):

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  • 收稿日期:2014-04-17
  • 最后修改日期:2014-04-17
  • 录用日期:2014-04-17
  • 在线发布日期: 2014-04-17
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