Mini-Tn7 transposons for site-specific tagging of bacteria with fluorescent proteins
Mini-Tn7 transposons for site-specific tagging of bacteria with fluorescent proteins
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DOI：
10.1111/j.1462-2920.2004.00605.x
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发表时间：
2004-07-01
影响因子：
5.1
通讯作者：
Molin, S
中科院分区：
文献类型：
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作者：
Lambertsen, L;Sternberg, C;Molin, S
微型Tn7转座子系统是一种方便的定点标记细菌的工具，其中标记的DNA被插入到唯一的和中性的染色体位置。我们扩大了表达不同荧光蛋白(稳定和不稳定)的微型Tn7递送质粒的范围，这些荧光蛋白来自大肠杆菌lac来源的启动子P-A1/04/03，或生长速度依赖的大肠杆菌启动子P-rrnB P1。将微型Tn7转座子插入土壤细菌恶臭假单胞菌KT2440中进行检测。通过Southern blots和聚合酶链式反应(PCR)验证了标记的正确性和特异性。此外，通过生长竞争实验检测荧光蛋白表达对细胞生长速率的影响。
The mini-Tn7 transposon system is a convenient tool for site-specific tagging of bacteria in which the tagging DNA is inserted at a unique and neutral chromosomal site. We have expanded the panel of mini-Tn7 delivery plasmids expressing different fluorescent proteins (stable and unstable) from the Escherichia coli lac derived promoter, P-A1/04/03, or from the growth-rate-dependent Escherichia coli promoter P-rrnB P1. The mini-Tn7 transposons were inserted and tested in the soil bacterium, Pseudomonas putida KT2440. Successful and site-specific tagging was verified by Southern blots as well as by PCR. Furthermore, the effect of fluorescent protein expression on the cellular growth rate was tested by growth competition assays.