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首页> 外文会议>International Colloquium on Paratuberculosis >Comparative genomic analysis of Mycobacterium avium ss. paratuberculosis isolates obtained from multiple host species
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Comparative genomic analysis of Mycobacterium avium ss. paratuberculosis isolates obtained from multiple host species

机译：分枝杆菌SS的比较基因组分析。从多个宿主物种中获得的副毛细血管分离物

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We have designed and built a DNA microarray consisting of 70mer oligonucoeotides representing all of the ORFs identified in the MAP K10 genome sequence as well as intergenic regions in order to investigate whether the genome content of Mycobacterium avium subspecies paratuberculosis (MAP) K10 is representative of other bovine isolates as well as isolates from other species. In addition, oligonucleotides representing coding sequences from the Mycobacterium avium subspecies avium (MAA) 104 genome that are not present in the MAP K10 genome were also included on the microarray. Genomic DNA from MAP isolates was fluorescently labeled, mixed with alternately labeled MAP K10 DNA, and competitively hybridized on the MAP microarray. ORFs were classified as present or divergent based on the relative fluorescent intensities of the experimental samples compared to MAP K10 DNA. MAP isolates cultured from cattle, bison, sheep, goat, avian, and human sources were hybridized to the MAP microarray. Three deleted regions were observed in the genomes of three MAP isolates obtained from sheep. Additionally, four clusters of ORFs homologous to sequences in the MAA 104 genome were identified in three of the sheep isolates. One of these clusters encodes glycopeptidolipid biosynthesis enzymes which have not previously been identified in MAP. Differences in hybridization across many of the isolates examined were detected for the microarray targets representing the insertion sequence IS_MAP04, the major membrane protein encoded by MAP2121c. Several regions encoding proteins with unknown function suggesting that there may be variation in the sequence or copy number of these regions. One cattle isolate and one sheep isolate of MAP were found to contain a different genome content compared to the other isolates examined from these species.
机译：我们已经设计和构建了由70mer寡核苷酸组成的DNA微阵列，其代表地图K10基因组序列中鉴定的所有ORF以及基因组织，以研究分枝杆菌亚副亚种的基因组含量是否是其他的代表性牛分离物以及来自其他物种的分离物。另外，在微阵列上也包括代表来自地图K10基因组的分枝杆菌亚阶亚型亚亚级（MAA）104基因组的编码序列的寡核苷酸也包括在微阵列上。来自Map分离株的基因组DNA被荧光标记，与交替标记的MAP K10 DNA混合，并在地图微阵列上竞争地杂交。与Map K10 DNA相比，基于实验样品的相对荧光强度分类ORF，与实验样品的相对荧光强度分类。地图分离株培养的牛，野牛，羊，山羊，禽，人类来源与地图微阵列杂交。在从绵羊获得的三种地图分离物的基因组中观察到三个缺失的区域。另外，在三种绵羊分离株中鉴定了对MAA 104基因组中的序列同源的四种ORF簇。其中一个簇编码了先前未在地图中鉴定的糖肽脂素生物合成酶。针对代表插入序列IS_map04的微阵列靶标检测杂交中的许多分离株的差异，由MAP2121C编码的主要膜蛋白。编码具有未知功能的蛋白质的几个区域表明这些区域的序列或拷贝数可能有变化。与从这些物种中检查的其他分离物相比，发现一个牛分离物和一个绵羊分离物含有不同的基因组含量。

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