Analysis of Customer Behaviour in the Cash & Carry Wholesale

Import 04/11/2015Tato bakalářská práce se zabývá analýzou nákupního chování mezi dvěma segmenty - vietnamskými a českými zákazníky, v ostravské prodejně Makro. Hlavním cílem práce je najít a porovnat rozdílné chování mezi těmito dvěma segmenty, ale i jejich podobnosti při nákupu v prodejně. Dílčím cílem je zjistit, proč zákazníci nenakupují v ostravské prodejně Makro častěji.This bachelor work focuses on the analysis of shopping behavior between the two segments - the Vietnamese and Czech customers in Ostrava Makro. The main objective is to find and compare the different behavior between these two segments, but also their similarities when customers buying in a store. Partial aim is to find out why customers do not buy in Ostrava Makro more often.116 - Katedra marketingu a obchoduvelmi dobř

Ornithine carbamoyltransferase (OCTase) specific activity of strains <i>P. syringae</i> pv. phaseolicola NPS3121 and YNorf1P.

<p>The strains analyzed are described under their corresponding value bars; YNorf1P is a derivative of NPS3121 with <i>phtA</i> inactivated by site-directed mutagenesis. The small numbers under the bars represents the temperature at which expression was carried out: 1 indicates 18°C and 2 indicates 28°C. Error bars represent standard deviation from triplicate samples.</p

Participation of genes from the phaseolotoxin biosynthesis cluster in the expression of gene <i>argK</i> from <i>P. syringae</i> pv. phaseolicola NPS3121.

<p>A. Graphic representation of the phaseolotoxin biosynthesis cluster (Pht cluster) of <i>P. syringae</i> pv. phaseolicola NPS3121. The Pht cluster contains five transcriptional units, including two monocistronic (<i>argK</i> and <i>phtL</i>) and three polycistronic operons (<i>phtA</i>, <i>phtD</i> and <i>phtM</i>) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046815#pone.0046815-Aguilera1" target="_blank">[25]</a>. There is a secondary promoter (P_D) capable of driving the expression of genes from <i>phtD</i> to <i>phtK</i>. Genes are represented by arrows, with the direction of the arrow indicating the direction of transcription. B. Analysis of the <i>argK</i> transcriptional pattern in <i>P. syringae</i> pv. phaseolicola NPS3121 and polar mutants by reverse transcription-PCR. RT-PCR amplicons were separated by electrophoresis, and reversed images of the gels are shown; the strains analyzed and the corresponding mutated genes are described on top of their corresponding lanes. The small numbers under the lanes represent the temperature at which expression was assayed: 1 indicates 18°C and 2 indicates 28°C.</p

Schematic representation of plasmid clones used for expression analyses.

<p>Each arrow represents each gene amplified and cloned into pUCP20. Each plasmid name is indicated at left. The direction of the arrow indicates the direction of transcription. All restriction sites for PstI, SmaI, SalI, BamHI and EcoRI are shown. A. Constructs used in Northern blot analyses. B. Constructs to evaluate the activity of the <i>argK</i> gene using transcriptional <i>uidA</i> fusions.</p

Gel retardation assay using an <i>argK</i> promoter (P_K) probe. The strains analyzed are described on top of their corresponding lanes.

<p>A. Gel retardation and competition assays using <i>P. syringae</i> pv. phaseolicola NPS3121 and CYL233. B. Gel retardation assay using <i>E. coli</i> DH5α and <i>P. syringae</i> pv. glycinea PG4180, pv. syringae B728a and pv. tomato DC3000. The P_K probe plus extract from the wild type strain NPS3121 grown at 28°C was used as positive control. The extract from <i>E. coli</i> DH5α was obtained from cells grown in LB at 37°C. “a" and “b" indicate the position of retardation signals; “c" indicates the position of free probe. The numbers on top of each lane represent the temperatures at which the cells were grown: 1 indicates 18°C and 2 indicates 28°C.</p

Expression of transcriptional <i>uidA</i> reporter gene fusions in <i>P</i>. <i>syringae</i> pv. phaseolicola NPS3121 and mutant backgrounds.

<p>3121phtA and 3121phtD correspond to <i>phtA</i>^<i>-</i> and <i>phtD</i>^<i>-</i> polar mutants, whereas NPS3121 indicates the wild type strain. As negative control, NPS3121 harboring pRG960sd was used. (A) GUS activity from pPphtA::GUS, which corresponds to the promoter of <i>phtA</i> operon cloned into vector pRG960sd. (B) GUS activity from pPphtD::GUS, corresponding to the promoter of <i>phtD</i> operon cloned into pRG960sd. The small numbers under the bars represent the temperatures at which expression was assayed: 1 indicates 18°C and 2 indicates 28°C. Bars represent mean values with standard deviations; bars in each panel topped with different letters indicate means that are significantly different according to a two-way ANOVA (<i>P</i>< 0.01) followed by the Duncan’s test.</p

Effect of cloned <i>argK</i> and <i>phtABC</i> genes on the <i>argK</i> expression in <i>P. syringae</i> pv. phaseolicola.

<p>The expression of <i>argK</i> was evaluated by Northern blot in derivatives of strain CYL233 (nontoxigenic; panels A and B) and strain NPS3121 (toxigenic, panel C) harboring plasmids with diverse combinations of genes <i>argK</i>, <i>phtA</i>, <i>phtB</i> and <i>phtC</i>, as indicated above each blot (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046815#pone-0046815-g003" target="_blank">Figure 3</a> for a description of each plasmid). Panel D shows the expression of <i>argK</i> in the absence or presence of carbamoylphosphate (CP) in strains NPS3121 and CYL233 at 28°C. Blots were hybridized with an internal probe specific for <i>argK</i>, and the signal corresponding to the monocistronic <i>argK</i> RNA is marked. The asterisks indicate the position of a band of approximately 2.3-kb, corresponding to a previously described possible alternative <i>argK</i> transcript. Strain CYL233(pUCP20) was used as negative control of <i>argK</i> expression, whereas the wild type strain NPS3121 was used as a positive control. The numbers on top of the Northern blots represent the temperatures at which expression was assayed: 1 indicates 18°C and 2 indicates 28°C.</p

Phaseolotoxin production by <i>P</i>. <i>syringae</i> pv. phaseolicola strains.

<p>(A) Structure of phaseolotoxin (top), which is cleaved by plant peptidases (arrow) to release octicidin, with indication of the inorganic moiety (N’-sulfodiaminophosphinyl). Octicidin is a transition state analog, structurally similar to carbamoylphosphate and ornithine (bottom), substrates of OCTase during biosynthesis of citrulline. (B) Evaluation of the production of phaseolotoxin by the wild type strain NPS3121 and mutant 3121phtD using the <i>E</i>. <i>coli</i> growth inhibition assay.</p

Bacterial strains and plasmids.

<p>Bacterial strains and plasmids.</p

Reverse transcription-PCR of the Pht cluster of <i>P</i>. <i>syringae</i> pv. phaseolicola NPS3121 and mutants.

<p>(A) Graphic representation of the Pht cluster of <i>P</i>. <i>syringae</i> pv. phaseolicola NPS3121. Genes are represented by arrows that indicate the direction of transcription. The Pht cluster contains five transcriptional units, including two monocistronic (<i>argK</i> and <i>phtL</i>) and three polycistronic (<i>phtA</i>, <i>phtD</i> and <i>phtM)</i> [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0178441#pone.0178441.ref018" target="_blank">18</a>]. Mutated genes are indicated by an X over the corresponding gene. (B) Analysis by RT-PCR of the transcriptional pattern of the Pht-cluster in the wild type strain NPS3121 and derivative mutants. Pictures show the RT-PCR product corresponding to each analyzed locus, as indicated by the black bars, separated by electrophoresis on agarose gels; mutated genes are indicated by an X over the corresponding picture of the RT-PCR gel. Numbers under the pictures represent the temperature at which expression was assayed: 1 = 18°C; 2 = 28°C.</p