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Mutational analysis of cj0183 Campylobacter jejuni promoter.

Salamasznska-Guz A, Grodzik M, Klimuszko D - Curr. Microbiol. (2013)

Bottom Line: It encodes a protein which shows partial homology to TlyC of Brachyspira hyodysenteriae.The aim of this work was to determine the mechanisms of gene regulation by cloning DNA fragments lying upstream of the cj0183 gene.Mutations in cj0183 -10 region, -16 region, and -35 region resulted in changes in gene transcription.

View Article: PubMed Central - PubMed

Affiliation: Department of Pre-Clinical Sciences, Warsaw University of Life Sciences, Warsaw, Poland, asalam@tlen.pl.

ABSTRACT
Gene-nominated cj0183 was identified in Campylobacter jejuni NCTC 11168 and in two human isolates 81116 and 81-176. It encodes a protein which shows partial homology to TlyC of Brachyspira hyodysenteriae. The aim of this work was to determine the mechanisms of gene regulation by cloning DNA fragments lying upstream of the cj0183 gene. The β-galactosidase activity determined for the strain harboring the plasmid with the fragment upstream of cj0183 indicated the presence of a promoter in this DNA region. Mutations in cj0183 -10 region, -16 region, and -35 region resulted in changes in gene transcription.

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Mutational analysis of the cj0183 promoter region. Quantitative analysis of lacZ expression of the cj0183 promoter deletion/substitution vector series in C. jejuni, including the construct carrying the wild-type cj0183 promoter (WT) as a positive control and the empty vector (control) as a negative control. Reactions were performed in triplicate, and standard deviations are marked by error bars. *P > 0.05
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Fig3: Mutational analysis of the cj0183 promoter region. Quantitative analysis of lacZ expression of the cj0183 promoter deletion/substitution vector series in C. jejuni, including the construct carrying the wild-type cj0183 promoter (WT) as a positive control and the empty vector (control) as a negative control. Reactions were performed in triplicate, and standard deviations are marked by error bars. *P > 0.05

Mentions: We constructed various deletions mutants within the assumed promoter region by removing: (i) the whole promoter region (Δ-35-16-10), (ii) -16 and -35 motifs (Δ-35-16), or (iii) only the -35 motif (Δ-35) (Fig. 2). All of these deletions significantly affected the activity of the β-galactosidase reporter gene (Fig. 3). Deletion of the -35-16-10 and -35-16 region abolished completely and deletion of the -35 region decreased threefold β-galactosidase activity, indicating that these regions are involved in gene transcription under standard laboratory conditions.Fig. 2


Mutational analysis of cj0183 Campylobacter jejuni promoter.

Salamasznska-Guz A, Grodzik M, Klimuszko D - Curr. Microbiol. (2013)

Mutational analysis of the cj0183 promoter region. Quantitative analysis of lacZ expression of the cj0183 promoter deletion/substitution vector series in C. jejuni, including the construct carrying the wild-type cj0183 promoter (WT) as a positive control and the empty vector (control) as a negative control. Reactions were performed in triplicate, and standard deviations are marked by error bars. *P > 0.05
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3824568&req=5

Fig3: Mutational analysis of the cj0183 promoter region. Quantitative analysis of lacZ expression of the cj0183 promoter deletion/substitution vector series in C. jejuni, including the construct carrying the wild-type cj0183 promoter (WT) as a positive control and the empty vector (control) as a negative control. Reactions were performed in triplicate, and standard deviations are marked by error bars. *P > 0.05
Mentions: We constructed various deletions mutants within the assumed promoter region by removing: (i) the whole promoter region (Δ-35-16-10), (ii) -16 and -35 motifs (Δ-35-16), or (iii) only the -35 motif (Δ-35) (Fig. 2). All of these deletions significantly affected the activity of the β-galactosidase reporter gene (Fig. 3). Deletion of the -35-16-10 and -35-16 region abolished completely and deletion of the -35 region decreased threefold β-galactosidase activity, indicating that these regions are involved in gene transcription under standard laboratory conditions.Fig. 2

Bottom Line: It encodes a protein which shows partial homology to TlyC of Brachyspira hyodysenteriae.The aim of this work was to determine the mechanisms of gene regulation by cloning DNA fragments lying upstream of the cj0183 gene.Mutations in cj0183 -10 region, -16 region, and -35 region resulted in changes in gene transcription.

View Article: PubMed Central - PubMed

Affiliation: Department of Pre-Clinical Sciences, Warsaw University of Life Sciences, Warsaw, Poland, asalam@tlen.pl.

ABSTRACT
Gene-nominated cj0183 was identified in Campylobacter jejuni NCTC 11168 and in two human isolates 81116 and 81-176. It encodes a protein which shows partial homology to TlyC of Brachyspira hyodysenteriae. The aim of this work was to determine the mechanisms of gene regulation by cloning DNA fragments lying upstream of the cj0183 gene. The β-galactosidase activity determined for the strain harboring the plasmid with the fragment upstream of cj0183 indicated the presence of a promoter in this DNA region. Mutations in cj0183 -10 region, -16 region, and -35 region resulted in changes in gene transcription.

Show MeSH
Related in: MedlinePlus