...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Clinical Microbiology >Distribution of a Nocardia brasiliensisCatalase Gene Fragment in Members of the GeneraNocardia, Gordona, andRhodococcus
【24h】

Distribution of a Nocardia brasiliensisCatalase Gene Fragment in Members of the GeneraNocardia, Gordona, andRhodococcus

机译：巴西诺卡氏菌，戈多纳和红球菌属成员中的巴西诺卡氏菌过氧化氢酶基因片段的分布

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

An immunodominant protein from Nocardia brasiliensis, P61, was subjected to amino-terminal and internal sequence analysis. Three sequences of 22, 17, and 38 residues, respectively, were obtained and compared with the protein database from GenBank by using the BLAST system. The sequences showed homology to some eukaryotic catalases and to a bromoperoxidase-catalase from Streptomyces violaceus. Its identity as a catalase was confirmed by analysis of its enzymatic activity on H2O2 and by a double-staining method on a nondenaturing polyacrylamide gel with 3,3′-diaminobenzidine and ferricyanide; the result showed only catalase activity, but no peroxidase. By using one of the internal amino acid sequences and a consensus catalase motif (VGNNTP), we were able to design a PCR assay that generated a 500-bp PCR product. The amplicon was analyzed, and the nucleotide sequence was compared to the GenBank database with the observation of high homology to other bacterial and eukaryotic catalases. A PCR assay based on this target sequence was performed with primers NB10 and NB11 to confirm the presence of the NB10-NB11 gene fragment in several N. brasiliensis strains isolated from mycetoma. The same assay was used to determine whether there were homologous sequences in several type strains from the generaNocardia, Rhodococcus, Gordona, andStreptomyces. All of the N. brasiliensisstrains presented a positive result but only some of the actinomycetes species tested were positive in the PCR assay. In order to confirm these findings, genomic DNA was subjected to Southern blot analysis. A 1.7-kbp band was observed in the N. brasiliensis strains, and bands of different molecular weight were observed in cross-reacting actinomycetes. Sequence analysis of the amplicons of selected actinomycetes showed high homology in this catalase fragment, thus demonstrating that this protein is highly conserved in this group of bacteria.
机译：来自巴西诺卡氏菌的一种免疫优势蛋白，P61，经过氨基末端和内部序列分析。获得了三个分别为22、17和38个残基的序列，并使用BLAST系统与GenBank中的蛋白质数据库进行了比较。这些序列与某些真核过氧化氢酶和紫链霉菌的溴过氧化物酶-过氧化氢酶具有同源性。通过分析其对H 2 O 2 的酶活性并在具有3,3'-的非变性聚丙烯酰胺凝胶上采用双重染色方法，证实了其为过氧化氢酶的身份。二氨基联苯胺和铁氰化物;结果仅显示过氧化氢酶活性，而没有过氧化物酶。通过使用内部氨基酸序列之一和共有过氧化氢酶基序（VGNNTP），我们能够设计产生500 bp PCR产物的PCR分析。分析扩增子，并将核苷酸序列与GenBank数据库进行比较，观察到与其他细菌和真核过氧化氢酶的高度同源性。用引物NB10和NB11进行基于该靶序列的PCR分析，以确认NB10-NB11基因片段在几个N中的存在。从菌丝瘤中分离出的巴西利亚菌株。使用相同的测定法确定来自诺卡氏菌，红球菌，戈多纳和的几种类型菌株中是否存在同源序列链霉菌。所有 N。巴西利亚菌株呈现阳性结果，但仅部分放线菌属物种在PCR分析中呈阳性。为了证实这些发现，对基因组DNA进行了Southern印迹分析。在 N中观察到1.7-kbp带。在交叉反应放线菌中观察到了巴西利亚菌株和不同分子量的条带。所选放线菌的扩增子的序列分析表明该过氧化氢酶片段具有高度同源性，因此证明该蛋白在这一组细菌中高度保守。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号