科微学术

微生物学通报

利用转座质粒plasposon构建荧光标记的脱色希瓦氏菌S12
DOI:
CSTR:
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金(No.30670020),国家高技术研究发展计划(863)项目(No.2006AA06Z322),广东省科技攻关项目(No.2004A30404002)


Construction of Fluorescent Labeled Shewanella decolorationis S12 Using Plasposon
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    采用分子生物学手段将具有转座功能的自杀性质粒pTnMod-okm与荧光蛋白基因eyfp构建重组质粒pTE-okm。pTE-okm通过结合转移进入脱色希瓦氏菌S12中,质粒上的转座子元件转座到S12的染色体上,而质粒本身的窄宿主复制位点使其在S12中不能得到有效的复制而“自杀”。荧光显微镜下筛选表达荧光蛋白的脱色希瓦氏菌克隆,通过对其提取质粒确定pTE-okm已经在脱色希瓦氏菌中自杀。筛选得到生长速度未发生延迟、脱色能力不受影响的荧光标记菌株S12-40。标记的脱色希瓦氏菌在无抗生素压力的情况下培养,传代20次(8h/次)后在荧光显微镜下依然查看到荧光蛋白的表达。该菌株的构建为研究其生态学行为奠定了基础。

    Abstract:

    The plasmid pTE-okm, which was obtained by inserting the fluorescence gene eyfp into the plasposon pTnMod-okm, transferred from E.coli S17-1 into Shewanella decolorationis S12 by conjugation. pTE-okm could not replicate in Shewanella decolorationis S12 because of its narrow-host-range replication origin, but the minitransposon within pTE-okm could transpose to the chromosome of Shewanella decolorationis S12 and made it survive the LB medium with kanamycin and rifampicin. The clones which expressed fluorescence gene eyfp were screened from all the clones on the selected plates under fluorescent microscope and it was make sure that pTE-okm didn't exist in these clones. S12-40, which had the the same growth rate and decoloration ability with the original strain S12-1, was picked out within these clones. S12-40 expressed EYFP stably even after successive re-culturing for 20 times (8h/time). The construction of S12-40 laid a foundation for the study on the ecological performance of Shewanella decolorationis S12.

    参考文献
    相似文献
    引证文献
引用本文

李光飞,邓代永,许枚英,岑英华,孙国萍. 利用转座质粒plasposon构建荧光标记的脱色希瓦氏菌S12[J]. 微生物学通报, 2007, 34(6): 1174-1178

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期:
  • 出版日期:
文章二维码