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1.
Fig. 7

Fig. 7. From: Transcription and genetic analyses of a putative N-acetylmuramyl-L-alanine amidase in Borrelia burgdorferi.

Phenotypic analysis of Δbb0666. The middle stationary phase cultures of the wild type, Δbb0666 and the complemented strain Δbb0666+ were observed by dark-field microscopy. At least 20 fields were examined for each strain.

Yu Yang, et al. FEMS Microbiol Lett. ;290(2):164-173.
2.
Fig. 6

Fig. 6. From: Transcription and genetic analyses of a putative N-acetylmuramyl-L-alanine amidase in Borrelia burgdorferi.

Characterization of Δbb0666 and the complemented strain Δbb0666+ by Western blot analysis. Approximately 10 μg of whole cell lysates were run by SDS-PAGE, and the polyclonal antiserum against Borrelia burgdorferi BB0666 was used for Western blots. WT, wild type.

Yu Yang, et al. FEMS Microbiol Lett. ;290(2):164-173.
3.
Fig. 1

Fig. 1. From: Transcription and genetic analyses of a putative N-acetylmuramyl-L-alanine amidase in Borrelia burgdorferi.

Constructing plasmids for the inactivation of bb0666 and the complementation of ΔBBB066. (a) The plasmid pbb0666∷kan was used to inactivate bb0666. (b) The plasmid FlgB666/pKFSS1 was used to complement Δbb0666. Arrows indicate the relative positions of the primers used for constructing these plasmids. The sequences of these primers are listed in . The symbol `Δ'shows that a 96-bp HindIII/MunI fragment within bb0666 was deleted and replaced with the kan cassette.

Yu Yang, et al. FEMS Microbiol Lett. ;290(2):164-173.
4.
Fig. 3

Fig. 3. From: Transcription and genetic analyses of a putative N-acetylmuramyl-L-alanine amidase in Borrelia burgdorferi.

Characterization of the pami operon. (a) The diagram showing the genes upstream of the flaA operon. The numbers and small arrows show the primers used for RT-PCR analysis. The relative positions of the promoters Pami and PflaA are labeled. (b) RT-PCR analysis. The numbered RT-PCR products were correspondingly produced by the primers labeled in (a). The sequences of these primers are described in . M, 1-kb DNA ladder.

Yu Yang, et al. FEMS Microbiol Lett. ;290(2):164-173.
5.
Fig. 4

Fig. 4. From: Transcription and genetic analyses of a putative N-acetylmuramyl-L-alanine amidase in Borrelia burgdorferi.

The pami operon is initiated by a σ70-like promoter, Pami. (a) Primer extension assay. The DNA ladder was generated with a DNA-sequencing reaction using Borrelia burgdorferi flaA gene as a template. The number is the size of the cDNA product generated from the primer extension assay. (b) Comparison of the Pami sequence with other σ70-like promoters identified in Escherichia coli and B. burgdorferi (, ). The asterisk indicates transcriptional initiation sites.

Yu Yang, et al. FEMS Microbiol Lett. ;290(2):164-173.
6.
Fig. 2

Fig. 2. From: Transcription and genetic analyses of a putative N-acetylmuramyl-L-alanine amidase in Borrelia burgdorferi.

Multiple-sequence alignment of the catalytic domains of amidases. Residues with 100% identity are shaded in black. GenBank accession numbers for the aligned sequences are as follows: Bacillus subtilis CwlC (BAA03500), Borrelia burgdorferi BB0666 (NP_212800), Treponema pallidum AmiA (NP_218687), Chlamydia pneumoniae AmiA (NP_224617), Escherichia coli AmiB (NP_290799), Salmonella typhimurium AmiB (NP_463219), Neisseria meningitidis AmiC (AF194079) and Helicobacter pylori AmiA (NP_207565). The residues labeled with * and+have been studied by amino acid substitutions in CwlC. *Essential residues for the catalytic activity of CwlC (). The alignments were performed using the program ALIGNX (Invitrogen).

Yu Yang, et al. FEMS Microbiol Lett. ;290(2):164-173.
7.
Fig. 5

Fig. 5. From: Transcription and genetic analyses of a putative N-acetylmuramyl-L-alanine amidase in Borrelia burgdorferi.

Transcriptional analysis of PflaA, Pami, PflgB and PflaB promoters using gfp as a reporter. The levels of GFP in (a) Escherichia coli and (b) Borrelia burgdorferi were detected by Western blot analysis. Approximately 10 μg of whole cell lysates were run by SDS-PAGE. The flagella motor switch complex protein, FliG, and the flagella filament protein, FlaB, were used as internal controls in E. coli and B. burgdorferi, respectively. The monoclonal antibodies against GFP and FlaB, and polyclonal antiserum against E. coli FliG, were used. (c) Quantitative Western blot analysis of GFP levels in E. coli and B. burgdorferi. Protein density was determined by FluorChem spot densitometry (Bio-Rad). Data were expressed as mean of density unit±SD of the mean from three independent experiments.

Yu Yang, et al. FEMS Microbiol Lett. ;290(2):164-173.

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