牛支原体不同分离株全菌蛋白免疫印迹分析

Abstract

Abstract: The whole proteins of 6 Mycoplasma bovis (M.bovis) isolates (W70 isolate,1738 isolate,Q3 isolate,Q1 isolate,JX02 isolate and 677 isolate),which isolated from different parts of the country was extracted by lysate,were analysed by SDS-PAGE and Western blotting with homemade antiserum.The results showed that there were differences on polypeptide quantity and definition of the whole proteins of 6 isolates:W70 isolate,1738 isolate,Q3 isolate and Q1 isolate were better than JX02 isolate and 677 isolate,which molecular weight were 23.2 to 130.8 ku.6 isolates were appeared 55 and 43 ku band when analysed by Western blotting.In conclusion,there were differences on polypeptide composition of whole proteins of the different M.bovis isolates,and 55 and 43 ku were the main immunogenicity protein of M.bovis.These results provided the theoretical basis for serology diagnosis,molecular diagnosis and the vaccine development of the M.bovis disease.

Key words: Mycoplasma bovis; whole proteins; SDS-PAGE; Western blotting

CLC Number:

S852.62

CHEN Zhong-qiong, FAN Yuan, JIANG Yong, ZHOU Bei, LI Xiao-min. Western Blotting Analysis on the Whole Proteins of Different Isolates of Mycoplasma bovis[J]. , 2014, 41(9): 59-62.

References

1 孔意端,陈杖榴,陈继荣,等.临床分离鸡毒支原体结构蛋白比较及抗原性分析[J].塔里木大学学报,2008,20(3):35～39.

2 孙文静,李媛,宋志强,等.牛支原体免疫相关性蛋白硫锌酰胺脱氢酶的筛选与鉴定[J].中国预防兽医学报,2012,34(10):761～765.

3 许健,储岳峰,高鹏程,等.我国17株绵羊肺炎支原体分子分型及其菌体蛋白免疫印迹[J].微生物学报,2011,51(10):1421～1426.

4 李媛,陈维,呼太飞,等.牛霉形体免疫相关蛋白P51的筛选和鉴定[J].中国兽医科学,2011,5:490～495.

5 萨姆布鲁克 J,拉塞尔 D W.分子克隆实验指南[M].第3版.北京:科学出版社,2002.

6 焦寒伟,王洪梅,刘晓,等.中国荷斯坦奶牛髓样分化蛋白-2 cDNA的克隆与原核表达[J].中国畜牧兽医,2012,39(2):20～23.

7 Gorg A,Obermaier C,Boguth G,et al.The current state of two-dimensional electrophoresis with immobilized pH gradients[J].Electrophoresis,2000,21(6):1037～1053.

8 Kumar A,Srivastava N C,Singh V P,et al.Antigenic analysis of M.bovis by SDS-PAGE and immunoblotting[A].Proceedings of the 20th Annual Conference of Indian Association of Veterinary Microbiologist and Immunologist,Pantnagar,India[C].2000.

9 Maid R,Patricia A,Semso P,et al.Protein and antigenic profile among Mycoplasma bovis field strains isolated in Bosnia and Herzegovina[J].Acta Vet,2009,78:151～154.

10 Nicholas R A J,Ayling R D.Mycoplasma bovis:Disease,diagnosis,and control[J].Research in Veterinary Science,2003,741(2):105～112.

11 Pfutzner H,Sachse K.Mycoplasma bovis as an agent of mastitis,pneumonia,arthritis and genital disorders [J].Scientific and Technical Review,1996,15(4):1477～1494.

12 Rabilloud T,Adessi C.Improvement of the solubilization of proteins in two-dimensional electrophoresis with immobilized pH gradients[J].Electrophoresis,1997,18:307～316.

13 Sachse K,Grajetzki C,Pfutzner H,et al.Comparison of Mycoplasma bovis strains based on SDS-PAGE and immunoblot protein patterns[J].Journal of Veterinary Medicine,1992,39(4):246～252.

14 Wilkins M R,Jones R C,Henry R R,et al.From proteins to proteomes:Large scale protein identification by two-dimensional electrophoresis and amino acid analysis[J].Biotechnology,1996,14:61～65.

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Western Blotting Analysis on the Whole Proteins of Different Isolates of Mycoplasma bovis

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