Characterization of cis-acting elements residing in the chitinase promoter of Bacillus pumilus SG2

Heravi, Kambiz Morabbi; Shali, Abbas; Naghibzadeh, N; Ahmadian, Gholamreza

doi:10.1007/s11274-013-1569-9

Cited by 10 publications

(9 citation statements)

References 33 publications

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“…A search for the B. pumilus cre (WTGNAARCGNWWWCA) consensus [81] indicated a potential site (TTGAAAACGAATTCA) located between two ORFs (14,389–14,403 bp) on contig 33 on which the bacilysin-like pathway was identified with a beta-glucoside transporter located 7 kb downstream on the same contig. Bacilysin production in Bacillus subtilis is however not subject to glucose repression [82].…”

Section: Resultsmentioning

confidence: 99%

Antibacterial Activities of Bacteria Isolated from the Marine Sponges Isodictya compressa and Higginsia bidentifera Collected from Algoa Bay, South Africa

et al. 2017

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Due to the rise in multi-drug resistant pathogens and other diseases, there is renewed interest in marine sponge endosymbionts as a rich source of natural products (NPs). The South African marine environment is rich in marine biota that remains largely unexplored and may represent an important source for the discovery of novel NPs. We first investigated the bacterial diversity associated with five South African marine sponges, whose microbial populations had not previously been investigated, and select the two sponges (Isodictya compressa and Higginsia bidentifera) with highest species richness to culture bacteria. By employing 33 different growth conditions 415 sponge-associated bacterial isolates were cultured and screened for antibacterial activity. Thirty-five isolates showed antibacterial activity, twelve of which exhibited activity against the multi-drug resistant Escherichia coli 1699, implying that some of the bioactive compounds could be novel. Genome sequencing of two of these isolates confirmed that they harbour uncharacterized biosynthetic pathways that may encode novel chemical structures.

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Section: Resultsmentioning

confidence: 99%

Antibacterial Activities of Bacteria Isolated from the Marine Sponges Isodictya compressa and Higginsia bidentifera Collected from Algoa Bay, South Africa

et al. 2017

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“…Elucidation of the inducing signal for YvoA Bt will require additional experiments. Heravi et al speculated that the chitinase gene (chiS) of Bacillus pumilus is under the control of CCR (36). Generally speaking, in low-GC Gram-positive bacteria such as B. subtilis, the key regulator for exerting CCR is CcpA.…”

Section: Discussionmentioning

confidence: 99%

YvoA and CcpA Repress the Expression of chiB in Bacillus thuringiensis

Jiang

Pan

et al. 2015

Appl Environ Microbiol

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c Bacillus thuringiensis produces chitinases, which are involved in its antifungal activity and facilitate its insecticidal activity. In our recent work, we found that a 16-bp sequence, dre chiB (AGACTTCGTGATGTCT), downstream of the minimal promoter region of the chitinase B gene (chiB) was a critical site for the inducible expression of chiB in B. thuringiensis Bti75. In this work, we show that a GntR family transcriptional regulator (named YvoA Bt ), which is homologous to YvoA of Bacillus subtilis, can specifically bind to the dre chiB oligonucleotide sequences in vitro by using electrophoretic mobility shift assays (EMSAs) and isothermal titration calorimetry (ITC) assays. The results of quantitative real-time reverse transcription-PCR (qRT-PCR) and Western blotting indicated that deletion of yvoA caused an ϳ7.5-fold increase in the expression level of chiB. Furthermore, binding of purified YvoA Bt to its target DNA could be abolished by glucosamine-6-phosphate (GlcN-6-P). We also confirmed, in the presence of the phosphoprotein Hpr-Ser 45 -P, that purified CcpA Bt bound specifically to the promoter of chiB, which contains the "cre chiB " sequence (ATAAAGCGTTTACA). According to the results of qRT-PCR and Western blotting, deletion of ccpA resulted in a 39-fold increase in the chiB expression level, and glucose no longer influenced the expression of chiB. We confirm that chiB is negatively controlled by both CcpA Bt and YvoA Bt in Bti75.

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“…PCR amplicons were cloned into the shuttle vector pDHAFB between the Hind III and Sph I restriction sites. To build pUPChi-CFP3 (lacking CRE box+sigma binding site), PCR1 was performed with the primers ChiSLF10 and UP-sig2 and PCR2 with primers ChiSR1J and UP-sig1, both using pUPChiΔcre[9] as a template (~700 and ~780 bp, respectively). These two PCR products were combined in equal volumes, and overlapping PCR was carried out using primers ChiSLF10 and ChiSR1J (~1480 bp).…”

Section: Methodsmentioning

confidence: 99%

“…The activity of this promoter was evaluated in B. subtilis as a host organism. The full-length ChiS promoter has been shown to have a limited function in protein expression; However, deletion of direct repeats or catabolite responsive elements (CRE) in the promoter leads to its increased activity[9,10]. Additionally, improved cyan fluorescent protein (ICFP) is a monochromic, green fluorescent protein (GFP) derivative produced by Aequorea macrodactyla in a process similar to that of GFP[11-15].…”

Section: Introductionmentioning

confidence: 99%

Expression and Secretion of Cyan Fluorescent Protein (CFP) in B. subtilis using the Chitinase Promoter from Bacillus pumilus SG2

Shali

Rigi²,

Pornour

et al. 2017

IBJ

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Background:Improved cyan fluorescent protein (ICFP) is a monochromic, green fluorescent protein (GFP) derivative produced by Aequorea macrodactyla in a process similar to GFP. This protein has strong absorption spectra at wavelengths 426-446 nm. ICFP can be used in cell, organelle or intracellular protein labeling, investigating the protein-protein interactions as well as assessing the promoter activities.Methods:In our previous study, the promoters of two chitinases (ChiS and ChiL) from Bacillus pumilus SG2 were assessed in B. subtilis and their regulatory elements were characterized. In the present study, icfp was cloned downstream of several truncated promoters obtained in the former study, and ICFP expression was evaluated in B. subtilis.Results:Extracellular expression and secretion of ICFP were analyzed under the control of different truncated versions of ChiSL promoters grown on different media. Results from SDS-PAGE and fluorimetric analyses showed that there were different expression rates of CFP; however, the UPChi-ICFP3 construct exhibited a higher level of expression and secretion in the culture medium.Conclusion:Our presented results revealed that inserting this truncated form of Chi promoter upstream of the ICFP, as a reporter gene, in B. subtilis led to an approximately ten fold increase in ICFP expression.

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Characterization of cis-acting elements residing in the chitinase promoter of Bacillus pumilus SG2

Cited by 10 publications

References 33 publications

Antibacterial Activities of Bacteria Isolated from the Marine Sponges Isodictya compressa and Higginsia bidentifera Collected from Algoa Bay, South Africa

Antibacterial Activities of Bacteria Isolated from the Marine Sponges Isodictya compressa and Higginsia bidentifera Collected from Algoa Bay, South Africa

YvoA and CcpA Repress the Expression of chiB in Bacillus thuringiensis

Expression and Secretion of Cyan Fluorescent Protein (CFP) in B. subtilis using the Chitinase Promoter from Bacillus pumilus SG2

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