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猪肺炎支原体田间菌株的基于芯片的基因组比较杂交分析

Array-based genomic comparative hybridization analysis of field strains of Mycoplasma hyopneumoniae.

作者信息

Madsen Melissa L, Oneal Michael J, Gardner Stuart W, Strait Erin L, Nettleton Dan, Thacker Eileen L, Minion F Chris

机构信息

Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, IA 50011, USA.

出版信息

J Bacteriol. 2007 Nov;189(22):7977-82. doi: 10.1128/JB.01068-07. Epub 2007 Sep 14.

Abstract

Mycoplasma hyopneumoniae is the causative agent of porcine enzootic pneumonia and a major factor in the porcine respiratory disease complex. A clear understanding of the mechanisms of pathogenesis does not exist, although it is clear that M. hyopneumoniae adheres to porcine ciliated epithelium by action of a protein called P97. Previous studies have shown variation in the gene encoding the P97 cilium adhesin in different strains of M. hyopneumoniae, but the extent of genetic variation among field strains across the genome is not known. Since M. hyopneumoniae is a worldwide problem, it is reasonable to expect that a wide range of genetic variability may exist given all of the different breeds and housing conditions. This variation may impact the overall virulence of a single strain. Using microarray technology, this study examined the potential variation of 14 field strains compared to strain 232, on which the array was based. Genomic DNA was obtained, amplified with TempliPhi, and labeled indirectly with Alexa dyes. After genomic hybridization, the arrays were scanned and data were analyzed using a linear statistical model. The results indicated that genetic variation could be detected in all 14 field strains but across different loci, suggesting that variation occurs throughout the genome. Fifty-nine percent of the variable loci were hypothetical genes. Twenty-two percent of the lipoprotein genes showed variation in at least one field strain. A permutation test identified a location in the M. hyopneumoniae genome where there is spatial clustering of variability between the field strains and strain 232.

摘要

猪肺炎支原体是猪地方流行性肺炎的病原体,也是猪呼吸道疾病综合征的主要因素。尽管很明显猪肺炎支原体通过一种名为P97的蛋白质作用粘附于猪的纤毛上皮,但对其发病机制尚无清晰的认识。先前的研究表明,不同猪肺炎支原体菌株中编码P97纤毛粘附素的基因存在变异,但不同地区分离株全基因组的遗传变异程度尚不清楚。由于猪肺炎支原体是一个全球性问题,考虑到所有不同的品种和饲养条件,预计可能存在广泛的遗传变异性。这种变异可能会影响单个菌株的整体毒力。本研究使用微阵列技术,检测了14株不同地区分离株与构建该微阵列所依据的232株相比的潜在变异情况。提取基因组DNA,用TempliPhi进行扩增,并用Alexa染料间接标记。基因组杂交后,扫描微阵列并使用线性统计模型分析数据。结果表明,在所有14株不同地区分离株中均能检测到遗传变异,但变异发生在不同的基因座上,这表明整个基因组都存在变异。59% 的可变基因座是假设基因。22% 的脂蛋白基因在至少一个不同地区分离株中显示出变异。一项排列检验确定了猪肺炎支原体基因组中的一个位置,在该位置不同地区分离株与232株之间的变异存在空间聚集。

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