Mini-Tn7载体作为一种遗传工具，用于在具有重要工业价值的植物病原细菌黄单胞菌属中以单拷贝数进行基因克隆。

Mini-Tn7 vectors as genetic tools for gene cloning at a single copy number in an industrially important and phytopathogenic bacteria, Xanthomonas spp.

作者信息

Jittawuttipoka Thichakorn, Buranajitpakorn Sarinya, Fuangthong Mayuree, Schweizer Herbert P, Vattanaviboon Paiboon, Mongkolsuk Skorn

机构信息

Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand.

出版信息

FEMS Microbiol Lett. 2009 Sep;298(1):111-7. doi: 10.1111/j.1574-6968.2009.01707.x. Epub 2009 Jun 26.

DOI:10.1111/j.1574-6968.2009.01707.x

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2782473/

Abstract

Transposon mini-Tn7 vectors insert into the chromosome of several Gram-negative bacteria in a site-specific manner. Here, we showed the application of mini-Tn7 as single copy site-specific integration vector system for Xanthomonas campestris pv. campestris. The transposition of the mini-Tn7 into the bacterial genome was detected at a Tn7 attachment (attTn7) site located downstream of glmS1. Furthermore, using a newly constructed vector pBBR1FLP2 containing the flipase (FLP) recombinase for site-specific excision of the sequence between the FLP recombinase target (FRT) sites, and a sacB counter selection marker, an unmarked mini-Tn7 insertion mutant was created. Mini-Tn7 insertion did not affect bacterial virulence on the tested plant. The mini-Tn7 and FLP-FRT systems also work well in Xanthomonas oryzae pv. oryzae.

摘要

转座子微型Tn7载体以位点特异性方式插入几种革兰氏阴性细菌的染色体中。在此，我们展示了微型Tn7作为野油菜黄单胞菌野油菜致病变种的单拷贝位点特异性整合载体系统的应用。微型Tn7向细菌基因组中的转座在位于glmS1下游的Tn7附着（attTn7）位点处被检测到。此外，使用新构建的含有翻转酶（FLP）重组酶以特异性切除FLP重组酶靶标（FRT）位点之间序列的载体pBBR1FLP2和一个sacB反选择标记，创建了一个无标记的微型Tn7插入突变体。微型Tn7插入不影响测试植物上细菌的毒力。微型Tn7和FLP-FRT系统在水稻黄单胞菌水稻致病变种中也能很好地发挥作用。