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发布于:2019-2-23 23:13:36  访问:19 次 回复:0 篇
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Ata). Our aim was to investigate {whether|whether or not|regardless
(2008) This function This workGeneral DNA methodsWhere essential, DNA restriction endonucleases, T4 DNA ligase, and alkaline phosphatase had been utilized as encouraged by the suppliers (NEB, New England Biolabs (Uk), Hitchin, UK).Motility and swarming assaysTo assess motility of Aeromonas strains, bacterial colonies had been transferred using a sterile toothpick into the center of motility agar plates (1 tryptone, 0.5 NaCl, 0.25 agar). The plates had been incubated face up at 25 for PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/22937147 14?four h, and motility was assessed by examining the migration of bacteria through the agar in the center toward the periphery on the plate. To assess the swarming capabilities of Aeromonas strains, bacterial colonies were transferred having a sterile toothpick in to the center of swarming agar plates (0.5 NaCl, 0.six Difco Nutrient Broth, and 0.6 Eiken agar). The plates have been incubated face up at 37 for 16 h, and swarming was assessed by examining the migration of bacteria orderAnacardic Acid across the agar in the center toward the periphery in the plate (Kirov et al.Ata). Our aim was to investigate irrespective of whether the AHA0618 gene solution
Ata). Our aim was to investigate regardless of whether the AHA0618 gene item could possibly be involved inside a doable deglycosylation step within the A. caviae flagellin glycosylation pathway that acts to modulate pseudaminic acid levels on its flagellin and regulate cellular behavior. Right here, we show that mutation of AHA0618 affects A. caviae swimming and swarming motility, but in spite of these behavioral changes, flagellin glycosylation levels don‘t appear to become altered in this bacterium.Components and MethodsBacterial strains, plasmids, and growth conditionsBacterial strains and PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/19502531 plasmids applied within this study are listed in Table 1. Escherichia coli strains have been grown in Luria?Bertani (LB) Miller broth and on LB Miller agar, though Aeromonas strains were grown in brain heart infusion broth (BHIB) or on Columbia blood agar (Oxoid, Basingstoke, UK). Development of E. coli and Aeromonas strains was commonly carried out at 37 . Ampicillin (50 lg/mL), nalidixic acid (50 lg/mL), kanamycin (50 lg/mL), gentamicin (25 lg/mL), streptomycin (50 lg/mL), and chloramphenicol (25 lg/mL) have been added when vital.?2014 The Authors. MicrobiologyOpen published by John Wiley Sons Ltd.AHA0618 Modulates Motility by Cell LengthR. C. Lowry et al.Table 1. Strains and plasmids employed in this study. Strain or plasmid Escherichia coli strains DH5a Genotype and use or description F?Phi80dlacZ DM15 D(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rK-mK+) phoA supE44 lambda-thi-1; applied for basic cloning hsdR pro recA, RP4-2 in chromosome, Km::Tn7 (Tc::Mu) kpir, Tpr Smr D(ara leu)7697 araD139 DlacX74 galE galK phoA20 thi-1 rspE rpoB(Rfr) argE(Am) recA1 kpir+ Sch, spontaneous Nalr Sch3N; AHA0618::kmr Cloning vector, Ampr Source of Tn5-derived nptII gene, Kmr oriR6K mobRK2 strAB sacBR, six.eight kb, Smr pBBR1MCS-5-derived broad-host-range expression vector containing lac promoter and lacIq, lacZa+, and Gmr pSRK(Gm) containing AHA0618 in NdeI/BamHI site of MCS pSRK(Gm) containing HP0518 in NdeI/BamHI web page of MCS Supply or referenceInvitrogenS17-1kpir CC118 kpir Aeromonas strains A. caviae Sch3N A. caviae JPS04 Plasmids pGEMT-EASY pUC4KIXX pKNG101 pSRK(Gm) pSRK_AHA0618 pSRK_HPde Lorenzo et al. (1990) Herrero et al.
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