全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

鹅细小病毒感染雏鹅的免疫球蛋白IgG/IgM变异性研究

DOI: 10.7668/hbnxb.2015.01.008, PP. 42-53

Keywords: 雏鹅,鹅细小病毒,免疫球蛋白M/G,SDS-PAGE,凝胶层析

Full-Text   Cite this paper   Add to My Lib

Abstract:

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)2SO4分级分离鹅血清Ig,将SephadexG-100凝胶柱在Utro-gelACA22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(128,114,69,59,55,48kDa);而在还原条件下,RD组缺失7个蛋白主带(139,128,119,113,97,94,61kDa),新增加3个蛋白主带(64,65,36kDa),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150kDa分子和重链(64kDa),而在还原条件下,鹅血清IgG显示重链(60~70kDa)特征蛋白带,重链62kDa特异蛋白带仅出现在CK组的IgM/G中,同时RD的IgG还缺失91kDa分子、IgG(ΔFc)(43kDa)、轻链(25kDa)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,62kDa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。

References

[1]  Zhao Q,Liu F,Chen S,et al.Chinese goose (Anser cygnoides) CD8a:cloning,tissue distribution and immunobiological in splenic mononuclear cells[J].Gene,2013,529(2):332-339.
[2]  Tsunekuni R,Ito H,Otsuki K,et al.Genetic comparisons between lentogenic Newcastle disease virus isolated from waterfowl and velogenic variants[J].Virus Genes,2010,40(2):252-255.
[3]  Sarker N,Tsudzuki M,Nishibori M,et al.Cell-mediated and humoral immunity and phagocytic ability in chicken Lines divergently selected for serum immunoglobulin M and G levels[J].Poultry Science,2000,79(12):1705-1709.
[4]  朱新产,王金宝,王宝维.鸡鸭鹅免疫球蛋白的研究进展[J].中国预防兽医学报,2003,25(6):503-508.
[5]  Guo Y,Gao M,Zhang H,et al.Molecular characterization and B cell membrane expression analysis of Fc fragment gene of goose IgY[J].Research in Veterinary Science,2014,pii:S0034-5288(14):00206-00209.
[6]  Yan X,Liu F,Chen S,et al.Molecular cloning,characterization and tissue expression of CD4 in Chinese goose[J].Gene,2013,519(2):298-304.
[7]  Huang L,Fang X.Immunoaffinity fractionation of plasma proteins by chicken IgY antibodies[J].Methods in Molecular Biology (Clifton,N.J.),2008,425:41-51.
[8]  张福淼,杨桂文,李国田,等.鲈血清免疫球蛋白分离纯化及部分特性分析[J].动物学杂志,2004,39(5):9-12.
[9]  蒋 卫,陈来同,胡晓倩,等.紫外分光光度法测定动物血清IgG含量方法的研究[J].中国生化药物杂志,2008,29(2):119-120.
[10]  夏 林,李荷华,董新善.免疫球蛋白测定方法的探讨及其临床应用[J].湖北医科大学学报,1994,15(2):161-164.
[11]  Warburg,Christian.Measuring protein concentration in the presence of nucleic acids by a280/a260:the method of warburg and christian[J].CSH Protoc,2006(1),pii:pdb.prot4252.
[12]  H?dge D,Ambrosius H.Comparative studies on the structure of biliary immunoglobulins of some avian species.I.Physico-chemical properties of biliary immunoglobulins of chicken,Turkey,duck and goose[J].Developmental and Comparative Immunology,1988,12(1):121-129.
[13]  Sharma J M.The structure and function of the avian immune system[J].Acta Veterinaria Hungarica,1997,45(3):229-238.
[14]  刘光磊,孙 健,王宝维,等.五龙鹅IgY、IgM分子特性研究[J].中国农业大学学报,2007,12(1):7-12.
[15]  Davies D R,Metzger H.Structural basis of antibody function[J].Annu Rev Immunol,1983,1:87-117.
[16]  Guo Y,Gao M,Wang J.A novel gene encoding goose immunoglobulin λ light chain[J].Acta Biochimica et Biophysica Sinica,2012,44(9):805-806.
[17]  Higgins D A,Warr G W.Duck immunoglobulins:structure,functions and molecular genetics[J].Avian Pathology,1993,22(2):211-236.
[18]  Humphrey B D,Calvert C C,Klasing K C.The ratio of full length IgY to truncated IgY in immune complexes affects macrophage phagocytosis and the acute phase response of mallard ducks (Anas platyrhynchos)[J].Developmental and Comparative Immunology,2004,28(7-8):665-672.
[19]  Law A J R,Leaver J.Effect of protein concentration on rates of thermal denaturation of whey protein in milk[J].J Agr ic Food chem,1997,45:4255-4261.
[20]  Mainer G,Sanchez L,Ena I M,et al.Kinetic and thermodynamix par ameters for heat denaturation of bovine milk I gG,IgA and IgM[J].J Food Sci,1997,62:1034-1038.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133