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To measure the speed of 89K excision, we produced an actual-time quantitative PCR (qPCR) assay and determined that the frequency of 89K excision from an 05ZYH33 overnight tradition was ∼0.032%, a rate similar to that noticed for SLG excision in the Streptomyces lividans sixty six
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The amplification specificity of every PCR was verified by pinpointing melting curves and by electrophoresis of the ultimate PCR products and solutions, and efficiency was firm applying linearized more info pMDph plasmid DNA, which is made up of a single copy of attP
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aspect, in addition to Other individuals derived from a wide variety of Gram-negative plant and animal pathogens, including Erwinia carotovora
Dependant on our experimental outcomes offered in this article, together with former scientific studies around the Agrobacterium
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The exact excision is mediated by an 89K-borne integrase via web site-precise recombination, with aid from an excisionase. The 89K excision intermediate acts as being a substrate for lateral transfer to non-89K S. suis
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