Acquisition of uncharacterized sequences from Candidatus Liberibacter, an unculturable bacterium, using an improved genomic walking method

文献类型： 外文期刊

第一作者： Doddapaneni, Harshavardhan

作者： Doddapaneni, Harshavardhan;Yao, Jiqiang;Civerolo, Edwin L.;Doddapaneni, Harshavardhan;Yao, Jiqiang;Bai, Xianjing;Zhao, Xiaolong

作者机构：

关键词： Genomic walking;Huanglongbing;Candidatus Liberibacter;Citrus greening

期刊名称：MOLECULAR AND CELLULAR PROBES （ 影响因子：2.365； 五年影响因子：2.386 ）

ISSN： 0890-8508

年卷期： 2008 年 22 卷 1 期

页码：

收录情况： SCI

摘要： An effective PCR-based genomic walking approach is described to discover previously unknown flanking genomic DNA sequences from Candidatus Liberibacter asiaticus, an unculturable, phloem-limited bacterium. Using this technique, 8564 bp of new DNA sequences were obtained from three genomic loci; tufB-secE-nusG-rplKAJL-rpoBC gene cluster, omp gene (outer membrane protein, Omp) and 16/23S rRNA gene in Ca. L. asiaticus. These, together with publicly available Ca. Liberibacter sequences, are clustered into five contigs and two singlets representing 24,477 non-redundant base pairs. BLAST annotation predicts 12 full-length genes, two partial genes and one pseudogene among these sequences. The sequences obtained in this study provide new genome information about Ca. Liberibacter that will facilitate development of new genome-based detection tools. The technique described here can also be employed to acquire new genomic information for other unculturable or fastidious organisms for which available sequences are limited or for filling sequence gaps between known flanking genomic DNA sequences. Published by Elsevier Ltd.

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