The ars Operon in the skinElement of Bacillus subtilis Confers Resistance to Arsenate and Arsenite

1998 ◽  
Vol 180 (7) ◽  
pp. 1655-1661 ◽  
Author(s):  
Tsutomu Sato ◽  
Yasuo Kobayashi
Keyword(s):  
Bacillus Subtilis ◽  
Constitutive Expression ◽  
Full Length ◽  
Primer Extension ◽  
Primer Extension Analysis ◽  
Negative Regulator ◽  
Arsenate Reductase ◽  
Northern Hybridization ◽  
Ars Operon ◽  
Extension Analysis

ABSTRACT The Bacillus subtilis skin element confers resistance to arsenate and arsenite. The ars operon in theskin element contains four genes in the orderarsR, ORF2, arsB, andarsC. Three of these genes are homologous to thearsR, arsB, and arsC genes from the staphylococcal plasmid pI258, while no homologs of ORF2have been found. Inactivation of arsR, arsB, orarsC results in either constitutive expression ofars, an arsenite- and arsenate-sensitive phenotype, or an arsenate-sensitive phenotype, respectively. These results suggest that ArsR, ArsB, and ArsC function as a negative regulator, a membrane-associated protein need for extrusion of arsenite, and arsenate reductase, respectively. Expression of the arsoperon was induced by arsenate, arsenite, and antimonite. Northern hybridization and primer extension analysis showed that synthesis of a full-length ars transcript of about 2.4 kb was induced by arsenate and that the ars promoter contains sequences that resemble the −10 and −35 regions of promoters that are recognized by EςA.

scholarly journals A muscle-specific intron enhancer required for rescue of indirect flight muscle and jump muscle function regulates Drosophila tropomyosin I gene expression.

1991 ◽  
Vol 11 (4) ◽  
pp. 1901-1911 ◽  
Author(s):  
J R Schultz ◽  
T Tansey ◽  
L Gremke ◽  
R V Storti
Keyword(s):  
Gene Expression ◽  
Flight Muscle ◽  
Primer Extension ◽  
Primer Extension Analysis ◽  
P Element ◽  
Indirect Flight Muscle ◽  
Mrna Levels ◽  
Enhancer Element ◽  
Mutant Phenotypes ◽  
Extension Analysis

The control of expression of the Drosophila melanogaster tropomyosin I (TmI) gene has been investigated by P-element transformation and rescue of the flightless and jumpless TmI mutant strain, Ifm(3)3. To localize cis-acting DNA sequences that control TmI gene expression, Ifm(3)3 flies were transformed with P-element plasmids containing various deletions and rearrangements of the TmI gene. The effects of these mutations on TmI gene expression were studied by analyzing both the extent of rescue of the Ifm(3)3 mutant phenotypes and determining TmI RNA levels in the transformed flies by primer extension analysis. The results of our analysis indicate that a region located within intron 1 of the gene is necessary and sufficient for directing muscle-specific TmI expression in the adult fly. This intron region has characteristics of a muscle regulatory enhancer element that can function in conjunction with the heterologous nonmuscle hsp70 promoter to promote rescue of the mutant phenotypes and to direct expression of an hsp70-Escherichia coli lacZ reporter gene in adult muscle. The enhancer can be subdivided further into two domains of activity based on primer extension analysis of TmI mRNA levels and on the rescue of mutant phenotypes. One of the intron domains is required for expression in the indirect flight muscle of the adult. The function of the second domain is unknown, but it could regulate the level of expression or be required for expression in other muscle.

Primer Extension Analysis of mRNA

2003 ◽  
pp. 195-199
Author(s):  
Maggie Walmsley ◽  
Mark Leonard ◽  
Roger Patient
Keyword(s):  
Primer Extension ◽  
Primer Extension Analysis ◽  
Extension Analysis

scholarly journals Multiplex primer extension analysis for rapid detection of major European mitochondrial haplogroups

2006 ◽  
Vol 27 (19) ◽  
pp. 3864-3868 ◽  
Author(s):  
Martina Wiesbauer ◽  
David Meierhofer ◽  
Johannes A. Mayr ◽  
Wolfgang Sperl ◽  
Bernhard Paulweber ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Primer Extension ◽  
Primer Extension Analysis ◽  
Mitochondrial Haplogroups ◽  
Extension Analysis

scholarly journals Organization of the 5′ region of the rat ATP citrate lyase gene

1994 ◽  
Vol 302 (3) ◽  
pp. 759-764 ◽  
Author(s):  
K S Kim ◽  
S W Park ◽  
Y A Moon ◽  
Y S Kim
Keyword(s):  
Genomic Library ◽  
Single Copy ◽  
Primer Extension ◽  
Primer Extension Analysis ◽  
Atp Citrate Lyase ◽  
Citrate Lyase ◽  
Sequence Elements ◽  
Flanking Region ◽  
Chloramphenicol Acetyltransferase Gene ◽  
Extension Analysis

A genomic clone, encompassing the 5′ flanking region and the first seven exons of rat ATP citrate lyase gene, was isolated from a rat genomic library and sequenced. Primer-extension analysis showed that mRNA is transcribed at 4407 nucleotides upstream from the translation start site. Primer-extension analysis and sequencing of ATP citrate lyase cDNA amplified by PCR showed that the promoter used for transcription is identical in mammary gland, lung, liver, brain and kidney. Southern-blot analysis showed that the ATP citrate lyase gene exists as a single copy. The 5′ flanking region contains several consensus sequences defined as promoter elements. These include a CAAT box and Sp1-binding sites. However, a TATA box lacks this promoter. The expression of the chloramphenicol acetyltransferase gene was induced by the 5‘ flanking region (-2370 to -1) in the CHO cell line. The 5′ flanking region also contains several sequence elements that may be involved in the transcriptional regulation of the gene.

scholarly journals Rapid detection of FGFR3 gene mutation in achondroplasia by DHPLC system-coupling heteroduplex and fluorescence-enhanced primer-extension analysis

2004 ◽  
Vol 49 (8) ◽  
pp. 399-403 ◽  
Author(s):  
Yi-Ning Su ◽  
Chien-Nan Lee ◽  
Shu-Chin Chien ◽  
Chia-Cheng Hung ◽  
Yin-Hsiu Chien ◽  
...  
Keyword(s):  
Gene Mutation ◽  
Rapid Detection ◽  
Primer Extension ◽  
Primer Extension Analysis ◽  
Fgfr3 Gene ◽  
System Coupling ◽  
Extension Analysis

scholarly journals T-box-mediated control of the anabolic proline biosynthetic genes of Bacillus subtilis

Microbiology ◽  
2011 ◽  
Vol 157 (4) ◽  
pp. 977-987 ◽  
Author(s):  
Jeanette Brill ◽  
Tamara Hoffmann ◽  
Harald Putzer ◽  
Erhard Bremer
Keyword(s):  
Protein Synthesis ◽  
Bacillus Subtilis ◽  
Northern Blot Analysis ◽  
Regulatory Mechanism ◽  
Full Length ◽  
Reporter Strain ◽  
Genetic Studies ◽  
T Box ◽  
Extension Analysis ◽  
Specific Control

Bacillus subtilis possesses interlinked routes for the synthesis of proline. The ProJ–ProA–ProH route is responsible for the production of proline as an osmoprotectant, and the ProB–ProA–ProI route provides proline for protein synthesis. We show here that the transcription of the anabolic proBA and proI genes is controlled in response to proline limitation via a T-box-mediated termination/antitermination regulatory mechanism, a tRNA-responsive riboswitch. Primer extension analysis revealed mRNA leader transcripts of 270 and 269 nt for the proBA and proI genes, respectively, both of which are synthesized from SigA-type promoters. These leader transcripts are predicted to fold into two mutually exclusive secondary mRNA structures, forming either a terminator or an antiterminator configuration. Northern blot analysis allowed the detection of both the leader and the full-length proBA and proI transcripts. Assessment of the level of the proBA transcripts revealed that the amount of the full-length mRNA species strongly increased in proline-starved cultures. Genetic studies with a proB–treA operon fusion reporter strain demonstrated that proBA transcription is sensitively tied to proline availability and is derepressed as soon as cellular starvation for proline sets in. Both the proBA and the proI leader sequences contain a CCU proline-specific specifier codon prone to interact with the corresponding uncharged proline-specific tRNA. By replacing the CCU proline specifier codon in the proBA T-box leader with UUC, a codon recognized by a Phe-specific tRNA, we were able to synthetically re-engineer the proline-specific control of proBA transcription to a control that was responsive to starvation for phenylalanine.

scholarly journals The ATP Synthase atpHAGDC(F1) Operon from Rhodobacter capsulatus

1998 ◽  
Vol 180 (2) ◽  
pp. 416-421 ◽  
Author(s):  
Roberto Borghese ◽  
Massimo Crimi ◽  
Luca Fava ◽  
Bruno Andrea Melandri
Keyword(s):  
Gene Transfer ◽  
Atp Synthase ◽  
Promoter Region ◽  
Rhodobacter Capsulatus ◽  
Growth Conditions ◽  
Primer Extension ◽  
Primer Extension Analysis ◽  
Viable Cells ◽  
Gene Transfer Agent ◽  
Extension Analysis

ABSTRACT The atpHAGDC operon of Rhodobacter capsulatus, containing the five genes coding for the F1 sector of the ATP synthase, has been cloned and sequenced. The promoter region has been defined by primer extension analysis. It was not possible to obtain viable cells carryingatp deletions in the R. capsulatus chromosome, indicating that genes coding for ATP synthase are essential, at least under the growth conditions tested. We were able to circumvent this problem by combining gene transfer agent transduction with conjugation. This method represents an easy way to construct strains carrying mutations in indispensable genes.

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