Abstract
Construction of gene disruption mutants and analysis of the resultant phenotypes are an important strategy to study gene function. A simple and high-throughput method developed for microorganisms combines two different types of transposons, direct genomic DNA amplification and thermal asymmetric interlaced-PCR. The considerable utility of this approach is demonstrable in Corynebacterium glutamicum, where 18,000 transposon disruptants enabled the generation of an insertion library covering nearly 80% of the organism’s 2,990 ORFs.
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Acknowledgments
We wish to thank Dr. C. Omumasaba for critical reading of the manuscript. This research was partly supported by New Energy and Industrial Technology Development Organization (NEDO), Japan.
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Suzuki, N., Inui, M., Yukawa, H. (2011). High-Throughput Transposon Mutagenesis of Corynebacterium glutamicum . In: Williams, J. (eds) Strain Engineering. Methods in Molecular Biology, vol 765. Humana Press. https://doi.org/10.1007/978-1-61779-197-0_24
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DOI: https://doi.org/10.1007/978-1-61779-197-0_24
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