Construction of targeted insertion mutations inFrancisella tularensissubsp.novicida
نویسندگان
چکیده
منابع مشابه
Construction of targeted insertion mutations in Francisella tularensis subsp. novicida.
Francisella tularensis is one of the most deadly bacterial agents, yet most of the genetic determinants of pathogenesis are still unknown. We have developed an efficient targeted mutagenesis strategy in the model organism F. tularensis subsp. novicida by utilizing universal priming of optimized antibiotic resistance cassettes and splicing by overlap extension (SOE). This process enables fast an...
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Taking advantage of the deep targeted sequencing capabilities of next generation sequencers, we have developed a novel two step insertion deletion (indel) detection algorithm (IDA) that can determine indels from single read sequences with high computational efficiency and sensitivity when indels are fractionally less compared to wild type reference sequence. First, it identifies candidate indel...
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Escherichia coli K12 strain x342LD requires two mutations in the bgl (@-glucosidase) operon, bglRo + bglR+ and excision of IS103 from within bglF, in order to utilize salicin. In growing cells the two mutations occur at rates of 4 X lo-* per cell division and <2 X 10”‘ per cell division, respectively. In 2-3-week-old colonies on MacConkey salicin plates the double mutants occur at frequencies o...
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Insertion mutagenesis via mobile genetic element is a common technique for the analysis of gene function in model organisms. Next-generation sequencing offers an attractive approach for localizing the site of insertion, but alignment-based mapping of mobile genetic elements is challenging. A computational method for identifying insertion sites is reported herein. The technique was validated by ...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 2007
ISSN: 0736-6205,1940-9818
DOI: 10.2144/000112574