Effective mutagenesis of Vibrio fischeri by using hyperactive mini-Tn5 derivatives.
نویسندگان
چکیده
We have developed a transposon mutagenesis system for Vibrio fischeri ES114 that utilizes a hyperactive mutant Tn5 transposase (E54K and M56A) and optimized transposon ends. Using a conjugation-based procedure, we obtained independent single-insertion mini-Tn5 mutants at a rate of approximately 10(-6). This simple and inexpensive technique represents a significant improvement over previous methods for transposon mutagenesis of V. fischeri and should be applicable to many other bacteria.
منابع مشابه
Transposon mutagenesis of marine Vibrio spp.
Coliphage P1 was used to transduce derivatives of transposons Tn5 and mini-Mu into marine Vibrio spp. Transposon Tn5 encoding tetracycline resistance (Tn5-132) was used to isolate mutants of Vibrio harveyi defective in genes for bioluminescence (lux). Insertion of transposon Tn5-132 into the lux gene region was demonstrated by intraspecific transduction with phage hv-1 and by Southern blot hybr...
متن کاملEngineering Vibrio fischeri for Inducible Gene Expression
The marine bacterium Vibrio fischeri serves as a model organism for a variety of natural phenomena, including symbiotic host colonization. The ease with which the V. fischeri genome can be manipulated contributes greatly to our ability to identify the factors involved in these phenomena. Here, we have adapted genetic tools for use in V. fischeri to promote our ability to conditionally control t...
متن کاملApplication of Vibrio qinghaiensis sp. Q67 for Ecotoxic Assessment of Environmental Waters – A Mini Review
Ecotoxicity assessment using luminescent bacteria has been wildly used because it is rapid, sensitive and cost effective for screening water and wastewater quality. This mini-review focused on the application of Vibrio qinghaiensis sp. Q67 (abbreviated as “Q67”), a natural freshwater luminescent bacteria strain discovered in China. The characteristics of the bioassay using Q67 were firstly revi...
متن کاملIn silico structural analysis of quorum sensing genes in Vibrio fischeri
Quorum sensing controls the luminescence of Vibrio fischeri through the transcriptional activator LuxR and the specific autoinducer signal produced by luxI. Amino acid sequences of these two genes were analyzed using bioinformatics tools. LuxI consists of 193 amino acids and appears to contain five α-helices and six ß-sheets when analyzed by SSpro8. LuxI belongs to the autoinducer synthetase fa...
متن کاملTransposon-induced non-motile mutants of Vibrio cholerae.
Non-motile mutants of Vibrio cholerae were isolated after transposon insertion mutagenesis with either Tn5 on a plasmid or Tn10ptac mini-kan in bacteriophage lambda. The physical location and number of transposon insertions was determined. Eighteen Tn5 insertion mutants and 11 Tn10ptac mini-kan insertion mutants had single unique insertion sites. The 18 Tn5 insertions were contained within six ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Applied and environmental microbiology
دوره 74 22 شماره
صفحات -
تاریخ انتشار 2008