鹅细小病毒感染雏鹅的免疫球蛋白IgG/IgM变异性研究  

Study on the Molecular Variability of Immunoglobulin IgG/IgM from Goslings Infected Goose Parvovirus

作　　者：朱新产[1] 邵艳红[1] 朱峰伟[1,2] 杨丽金[1] 

机构地区：[1]青岛农业大学生命科学学院山东省高校植物生物技术重点实验室,山东青岛266109 [2]青岛即墨畜牧兽医局,山东即墨266200

出　　处：《华北农学报》2015年第1期42-53,共12页Acta Agriculturae Boreali-Sinica

基　　金：山东省自然科学基金项目(ZR2011CM008);青岛市科技基金资助项目(12-1-4-5-(2)-jch);青岛市科技支撑计划项目(12-1-3-17-nsh)

摘　　要：为探究IgG／IgM蛋白的分子特征、理化特性及病理学相关致病机理，对鹅细小病毒感染雏鹅的免疫球蛋白Ig进行了分级分离、纯化及 IgG／IgM 蛋白质结构和功能等生化-分子生物学分析。以 GPVYZ 感染雏鹅，采用（ NH4）2 SO4分级分离鹅血清Ig，将Sephadex G-100凝胶柱在Utro-gel ACA22凝胶柱色谱工作站上串联，分步聚集纯化出鹅血清IgM／G进行SDS-PAGE和生化分析。结果显示，初级沉淀鹅血清Ig，在非还原条件下，感染组（ RD）缺失6个蛋白主带（128，114，69，59，55，48 kDa）；而在还原条件下，RD组缺失7个蛋白主带（139，128，119，113，97，94，61 kDa），新增加3个蛋白主带（64，65，36 kDa），RD组和对照组（CK）的IgM和IgG带分别增加10，7倍和1．5，2倍。表明鹅血清IgM、IgG单体的高度可变性造成了在形状构型上的多态性，其亚基条带的变化是机体抗病潜能的重要标志；纯化鹅血清IgM／G，在非还原条件下，RD组的IgG缺失150 kDa分子和重链（64 kDa），而在还原条件下，鹅血清IgG显示重链（60～70 kDa）特征蛋白带，重链62 kDa特异蛋白带仅出现在CK组的IgM／G中，同时RD的IgG还缺失91 kDa分子、IgG（ΔFc）（43 kDa）、轻链（25 kDa）蛋白带。提示GPV感染与鹅血清Ig发生相互作用，IgG与GPV感染密切相关，62 kDa重链基因可能是GPV的易感基因。试验还表明，RD组的鹅血清Ig含量仅为CK组的37％，IgG含量较IgM高2．56倍，RD组IgM、IgG比活力显著高于CK组，提示GPV拟制鹅血清IgM、IgG基因的表达，促使病毒感染雏鹅。稳定性试验显示，酸碱稳定性：鹅血清IgG＞IgM；热稳定性IgM＞IgG，RD组的IgM、IgG对碱和温度较为敏感。提示随pH值、温度的不同，Ig同时发生许多反应，GPV感染增加了鹅血清Ig的碱和温度敏感性。To conduct isolating and purifying of immunoglobulin IgG/IgM from goslings infected Goose parvo-virus YZ strain（GPV-YZ） and make analysis of protein structure and function by biochemical and molecular biolo-gy , and research the characteristics of Ig and the physicochemical properties and pathogenic mechanism of IgG /IgM protein.Immunoglobulin（Ig） was isolated and purified from goslings infected Goose parvovirus （RD）and control group（CK）by ammonium sulphate fractionation chromatographies on Sephadex G-100 and Utro-gel ACA22.The re-sults of SDS-PAGE and biochemical analysis showed that there were deleted 6 main protein（128,114,69,59,55,48 kDa ） band of the primary sedimentation Ig from serum of goslings infected Goose parvovirus in non reductive condi -tions,but under reductive conditions,deleted 7 main protein（139,128,119,113,97,94,61 kDa） band and ap-peared three new main protein （64,65,36 kDa） band of the primary sedimentation Ig from serum of goslings infec-ted Goose parvovirus .There were increased respectively 10 ,7 times or 1 .5 ,2 times of protein bands of IgM and IgG from goslings infected Goose parvovirus and control group , and indicated that the high variability macromolecular IgM/G monomers caused an abundant polymorphism in shape configuration ,and the IgM/G subunit change was an＆amp;nbsp;important marker of the resistance potential in goslings .There were deleted 150 kDa band and heavy chain （ 64 kDa） of the purified IgG from serum of goslings infected Goose parvovirus in non reductive conditions ,but under re-ductive conditions,the IgG showed heavy chain（60~70 kDa） characteristic protein band in goslings infected Goose parvovirus and control group .The 62 kDa specific heavy chain bands only appeared in the IgM/G of control group , and deleted the 91 kDa,IgG（ΔFc）（43 kDa）and light chain （25 kDa） protein band from serum of goslings infected Goose parvovirus .Those indicated that there existed the interaction between GPV infection and Ig .The IgG is as

关 键 词：雏鹅 鹅细小病毒 免疫球蛋白M/G SDS-PAGE 凝胶层析 

分 类 号：S853.5[农业科学—临床兽医学] Q78[农业科学—兽医学]