A gene coding for ornithine decarboxylase (odcA) is differentially expressed during the Mucor circinelloides yeast-to-hypha transition

Blasco, José L.; Garcı́a-Sánchez, Asunción; Ruíz-Herrera, José; Eslava, Arturo P.; Iturriaga, Enrique A.

doi:10.1016/s0923-2508(02)01301-3

Cited by 14 publications

(11 citation statements)

References 53 publications

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“…Normalization with the two most stably expressed genes identified for all stages, namely Ubc and WS21 , clearly demonstrated that the expression of Bp odc was lower in yeast cells, sporangiospores and zygospores as compared to the expression level obtained in hyphae, which is in accordance with the previous reports [14, 26–27]. However, normalization with Tub-b led to misinterpretation of the results and even a different conclusion.…”

Section: Discussionsupporting

confidence: 87%

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Selection of reference genes for quantitative real-time RT-PCR assays in different morphological forms of dimorphic zygomycetous fungus Benjaminiella poitrasii

2017

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Benjaminiella poitrasii, a dimorphic non-pathogenic zygomycetous fungus, exhibits a morphological yeast (Y) to hypha (H) reversible transition in the vegetative phase, sporangiospores (S) in the asexual phase and zygospores (Z) in the sexual phase. To study the gene expression across these diverse morphological forms, suitable reference genes are required. In the present study, 13 genes viz. ACT, 18S rRNA, eEF1α, eEF-Tu,eIF-1A, Tub-α, Tub-b, Ubc, GAPDH, Try, WS-21, NADGDH and NADPGDH were evaluated for their potential as a reference, particularly for studying gene expression during the Y-H reversible transition and also for other asexual and sexual life stages of B. poitrasii. Analysis of RT-qPCR data using geNorm, normFinder and BestKeeper software revealed that genes such as Ubc, 18S rRNA and WS-21 were expressed at constant levels in each given subset of RNA samples from all the morphological phases of B. poitrasii. Therefore, these reference genes can be used to elucidate the role of morpho-genes in B. poitrasii. Further, use of the two most stably expressed genes (Ubc and WS-21) to normalize the expression of the ornithine decarboxylase gene (Bpodc) in different morphological forms of B. poitrasii, generated more reliable results, indicating that our selection of reference genes was appropriate.

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Section: Discussionsupporting

confidence: 87%

“…The expression is usually higher in hyphae than in yeast cells [14, 26–27]. Therefore, to validate that we have made the correct choice of reference genes, RT-qPCR analysis was performed for Bp odc , Ubc , WS-21 and Tub-b .…”

Section: Resultsmentioning

confidence: 99%

Selection of reference genes for quantitative real-time RT-PCR assays in different morphological forms of dimorphic zygomycetous fungus Benjaminiella poitrasii

2017

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“…Primer extension and RT-PCR experiments indicate that the genes are transcribed as two messengers, one polycistronic (nasABC) and one monocistronic (nasD). This method for the determination of the transcription initiation sites has been successfully employed recently (Blasco et al, 2002). In two cases, initiation sites corresponded to those previously identified using the conventional radioactive procedure, demonstrating its accuracy (Altermann et al, 1999, Yamada et al, 1998.…”

Section: Discussionmentioning

confidence: 62%

Identification and transcriptional analysis of nitrate assimilation genes in the halophilic archaeon Haloferax mediterranei

Lledó

Marhuenda-Egea

Martínez‐Espinosa

et al. 2005

Gene

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“…To identify the transcription start site(s) of TLX3, we took advantage of a protocol for the automated detection of fluorescent primer extension products, adapted from the standard method based on radioactive labeling (18,19) and already successfully applied to other genes (20,21).…”

Section: Determination Of Transcriptional Start Sitesmentioning

confidence: 99%

“…With 10 Amol/L primer concentration, nonspecific background fluorescence occurred in the form of an increased baseline, thus suggesting 1 Amol/L as the most appropriate concentration for subsequent experiments. To obtain a precise determination of the transcriptional start point(s), we also run the primer extension reactions together with the DNA fragments generated in a sequencing reaction carried out with the same but unlabeled oligonucleotide (20). The peak detected 272 nucleotides upstream of the ''ATG'' translation start codon ( Fig.…”

Section: Determination Of Transcriptional Start Sitesmentioning

confidence: 99%

Nuclear Factor Y Drives Basal Transcription of the Human TLX3, a Gene Overexpressed in T-Cell Acute Lymphocytic Leukemia

Borghini

Vargiolu

Duca

et al. 2006

Molecular Cancer Research

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Based on a knocked-out mouse model and a few expression studies, TLX3 is regarded as a homeobox gene crucial for the development of the autonomic nervous system. This gene can undergo rearrangements or deregulation, giving rise to T-cell acute lymphocytic leukemia. The present report is focused on the identification of elements and factors playing a role in the TLX3 physiologic expression regulation and therefore likely to be involved in cancer development. In particular, after identifying the transcription start points, we have made use of in vitro transfection assays to show that the 5 ¶-untranslated region of the gene is necessary for the basal promoter activity in cell lines from different origin. By site-directed mutagenesis, two tandem CCAAT boxes have been localized as critical elements of this region. In vivo chromatin immunoprecipitation and electrophoretic mobility shift assays have indicated that nuclear factor Y (NFY) recognizes these sites in all the analyzed cell lines. The physiologic role of such an interaction has been confirmed by a dominant-negative version of the NFY transcription factor that has turned out to decrease both in vitro TLX3 promoter activity and endogenous amount of mRNA. Finally, a consistent in vivo TLX3 expression impairment was also achieved after NFY mRNA knockdown. The full characterization of the TLX3 transcription regulation will ultimately provide crucial elements to define the involvement of this gene in T-cell acute lymphocytic leukemia development. (Mol Cancer Res 2006;4(9):635 -43)

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A gene coding for ornithine decarboxylase (odcA) is differentially expressed during the Mucor circinelloides yeast-to-hypha transition

Cited by 14 publications

References 53 publications

Selection of reference genes for quantitative real-time RT-PCR assays in different morphological forms of dimorphic zygomycetous fungus Benjaminiella poitrasii

Selection of reference genes for quantitative real-time RT-PCR assays in different morphological forms of dimorphic zygomycetous fungus Benjaminiella poitrasii

Identification and transcriptional analysis of nitrate assimilation genes in the halophilic archaeon Haloferax mediterranei

Nuclear Factor Y Drives Basal Transcription of the Human TLX3, a Gene Overexpressed in T-Cell Acute Lymphocytic Leukemia

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