论文部分内容阅读
通过饱和硫酸铵分步盐析结合柱层析纯化温和气单胞菌免疫的斑点叉尾(鱼回)血清免疫球蛋白(Ig),结果表明,斑点叉尾(鱼回)血清Ig主要分布在35%-55%的硫酸铵沉淀区间;而在Sepharose-4B凝胶柱和DEAE-52阴离子交换柱层析纯化的Ig均出现在第1个蛋白质峰。SDS-PAGE分析表明,纯化后的血清Ig纯度提高。通过变性还原、变性非还原和非变性非还原3种条件下的免疫印迹(Western blot)试验对血清Ig的结构进行初步分析,发现SDS变性还原条件下血清Ig重链分子量为72 kD,轻链有3条.其分子量分别为21 kD、23.5 kD和26 kD;在变性非还原条件下血清Ig则出现多种结构形式,主要条带的分子量分别为760 kD、525 kD、330 kD和230 kD;而在非变性非还原条件下Ig仅出现一种结构形式,分子量约为870 kD。
The serum immunoglobulin (Ig) of Channel catfish (FISH) immunized with A. aeruginosa was purified by saturated ammonium sulfate fractionated salting-out combined with column chromatography. The results showed that the Ig of serum channel catfish 35% -55% ammonium sulphate precipitation interval; while in the Sepharose-4B gel column and DEAE-52 anion exchange column chromatography purified Ig are present in the first protein peak. SDS-PAGE analysis showed that purified serum Ig purity increased. The structure of serum Ig was analyzed preliminarily by denaturing, denaturing, non-denaturing and non-denaturing non-reducing conditions. The results showed that the molecular weight of serum Ig heavy chain was 72 kD and the light chain And its molecular weight is 21 kD, 23.5 kD and 26 kD, respectively. Under the denaturation and non-reducing conditions, there are many kinds of structural forms of Ig in serum, the main bands have molecular weights of 760 kD, 525 kD, 330 kD and 230 kD ; While in the non-denaturing non-reducing conditions, only one structure of Ig appeared, the molecular weight was about 870 kD.