摘要
通过DEAE Sepharose阴离子交换层析、SephacrylS 2 0 0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 .2 5 ,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 1 0 0 0 0 0g(2h)离心不沉降、耐热、在pH2 - 1 0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α/ β干扰素的特性相一致。
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 purified 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 NaIO 4. It was able to inhibit different kinds of virus in the cells of grass carp, but failed in human, mammalian, avian and shellfish cells. The results showed that the grass carp interferon was specific to fish cells and viruses. Its characterization was consistant with that of high vertibrate α/β interferon.
出处
《水产学报》
CAS
CSCD
北大核心
2000年第1期11-16,共6页
Journal of Fisheries of China
基金
国家自然科学基金!(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 )
3 940 0 10 0和 3 9970 5 89号
关键词
草鱼
干扰素
分离纯化
理化性质
生物学功能
Ctenocpharyngodon idellus, interferon, purification, Physico chemical characterization
