Transposon mutagenesis of Mycoplasma gallisepticum
Transposon mutagenesis of Mycoplasma gallisepticum
复制标题
DOI：
10.1016/s0147-619x(02)00114-2
复制
发表时间：
2003-01-01
期刊：
影响因子：
2.6
通讯作者：
Dohms, JE
中科院分区：
文献类型：
--
作者：
Whetzel, PL;Hnatow, LL;Dohms, JE
有几个系统可用于研究重要的禽呼吸道病原体，鸡毒支原体的遗传学。这些技术的发展需要一个机制来研究M的分子发病机制。鸡败血症Tn 916具有转座到M.通过转化和接合两种方式对鸡败血症病毒基因组进行修饰。本研究采用PEG介导法将Tn 916转化M.并创建转座子突变体文库。转化子的获得频率接近于5 × 10 ~（-8）/受体CFU。共构建了424个MG/Tn 916突变体，获得了71个突变体的转座子连接点的序列数据，并用于鉴定转座子插入位点。在几乎所有主要基因类别的基因组中都发现了插入，这使得这是M的转座子突变库的第一个广泛表征。鸡败血症转座子的稳定性也进行了检查，并确定，对于两个突变体的元件是稳定地保持在体内的选择压力的情况下。(C)2002 Elsevier Science（美国）。All rights reserved.
There are few systems available for studying the genetics of the important avian respiratory pathogen, Mycoplasma gallisepticum. These techniques are needed to develop a mechanism to study the molecular pathogenesis of M. gallisepticum. Tn916 has the ability to transpose into the M. gallisepticum genome by both transformation and conjugation. In this study, PEG-mediated transformation was employed for the transfer of Tn916 into M. gallisepticum and create a transposon mutant library. Transformants were obtained at a frequency of similar to5 x 10(-8) per recipient CFU. A total of 424 MG/Tn916 mutants were constructed and sequence data from the transposon junctions of 71 mutants was obtained and used to identify transposon insertion sites. Insertions were found throughout the genome in nearly all of the major gene categories, making this the first extensive characterization of a transposon mutant library of M. gallisepticum. Transposon stability was also examined, and it was determined that for two mutants the element was stably maintained in vivo in the absence of selective pressure. (C) 2002 Elsevier Science (USA). All rights reserved.