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鸭疫里氏杆菌吉林分离株16S rRNA系统分析及其OmpA基因克隆
高云航,马红霞
0
(吉林农业大学动物科技学院;吉林正方农牧股份有限公司)
摘要:
【目的】寻求RA 16S rRNA变异与OmpA基因之间的关系,可为进一步研究RA的变异及其亚单位疫苗提供一定参考依据。【方法】提取鸭疫里氏杆菌(RA)吉林分离株JL-RA1和JL-RA3总DNA,利用细菌通用引物扩增16S rRNA基因,并设计特异性引物扩增两株RA保护性抗原OmpA全基因序列。【结果】扩增出16S rRNA序列大小均为1478 bp,OmpA片段序列大小均为1164bp,与预期结果一致。【结论】扩增的16S rRNA与GenBank中已知的RA 16S rRNA序列进行同源性对比,同源性高达99.0%~99.9%。扩增的OmpA序列与GenBank中已知的RA OmpA序列进行同源性对比,同源性达93.3%~100%,编码蛋白质的氨基酸序列同源性为96%~100%。
关键词:  鸭疫里氏杆菌  16S rRNA基因  OmpA基因  系统进化分析
DOI:
投稿时间:2013-01-17修订日期:2013-04-11
基金项目:吉林省科技发展计划项目(20110221);国家现代农业产业技术体系建设专项(CARS-43)。
Systematic Analysis of 16S rRNA and Cloning of OmpA Gene of the Riemerella Anatipestifer
(College of Animal Science and Technology,Jilin Agricultural University;Jilin Zhengfang Agriculture and Animal Husbandry CoLtd)
Abstract:
【Objective】It is necessary to provide a reference basis for future study of the variation of RA and its subunit vaccine which seeking for a relationship between the variation of 16S rRNA of RA with the OmpA gene. 【Method】Extraction of total DNA of the Riemerella anatipestifer (RA) that were isolated from Jilin which named JL-RA1 and JL-RA3, the 16S rRNA sequence was amplified by bacterial universal primers,and both of the RA protective antigen OmpA gene were amplified by the specific primers. 【Result】The 16S rRNA sequences were 1478bp,and the OmpA sequences were 1164bp.【Conclusion】 Contrasting the homology of the RA 16S rRNA sequences with the sequences which was known in GenBank, the homology of 99.0% to 99.9%. and contrasting the homology of the OmpA sequences with the sequences of RA which was known in GenBank, the homology of 93.3% to 100%, and encoding a protein the amino acid sequence homology of 96% to 100%.
Key words:  Riemerella anatipestifer  16S rRNA gene  OmpA gene  Phylogenetic analysis

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