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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Transposon mutagenesis of Mycoplasma gallisepticum by conjugation with enterococcus faecalis and determination of insertion site by direct genomic sequencing.
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Transposon mutagenesis of Mycoplasma gallisepticum by conjugation with enterococcus faecalis and determination of insertion site by direct genomic sequencing.

机译：与粪肠球菌结合，对鸡败血支原体进行转座子诱变，并通过直接基因组测序确定插入位点。

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摘要

Few genetic systems for studying mycoplasmas exist, but transposon Tn916 has been shown to transpose into the genomes of some species and can be used as an insertional mutagen. In the current study, the ability of Enterococcus faecalis to serve as a donor for the conjugative transfer of transposon Tn916 into the genome of the avian pathogen Mycoplasma gallisepticum strain PG31 was examined. Transconjugants were obtained at a frequency of > or =6 x 10(-8) per recipient CFU. To determine the transposon insertion site, an oligonucleotide primer corresponding to the 3' end of Tn916 was designed for the purpose of directly sequencing genomic DNA without PCR amplification. Using the direct sequencing approach, Tn916 was shown to insert into any of numerous sites in the M. gallisepticum genome. This is the first report of conjugal transposition of Tn916 into the M. gallisepticum genome. The ability to determine transposon insertion sites in mycoplasmas by genomic sequencing has not been previously described and allows rapid sequence analysis of transposon-generated mutants. Copyright 2000 Academic Press.
机译：很少有用于研究支原体的遗传系统，但是转座子Tn916已被证明可转位到某些物种的基因组中，并可用作插入诱变剂。在当前的研究中，检查了粪肠球菌作为转座子Tn916转移到禽病原体支原体菌株PG31的基因组中的供体的能力。每个接受者CFU以>或= 6 x 10（-8）的频率获得转导结合体。为了确定转座子插入位点，设计了对应于Tn916 3'端的寡核苷酸引物，目的是直接对基因组DNA进行测序而不进行PCR扩增。使用直接测序方法，显示Tn916插入鸡毒支原体基因组的许多位点中的任何位点。这是首次将Tn916结合转座鸡毒支原体基因组的报道。以前没有描述过通过基因组测序确定支原体中转座子插入位点的能力，并允许对转座子产生的突变体进行快速序列分析。版权所有2000学术出版社。

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