Heterologous reporter genes are widely used for the characterization of gene expression in many organisms. Particularly, constructs bearing reporter genes have greatly contributed to our understanding of gene regulation in kinetoplastids. In some specific circumstances, however, such heterologous reporter has a risk of resulting in irrelevant observations and conclusions, which are primarily due to the introduction of foreign sequence elements. This communication describes our recent experience using the chloramphenicol acetyltransferase (CAT) gene as a reporter for analysis of the translational regulation of HSP70 genes in Leishmania infantum. We show that chimeric mRNAs consisting of the CAT open reading frame (ORF) and the untranslated regions (UTRs) from HSP70-II genes behave differently as endogenous HSP70-II mRNAs and that this difference is due to the presence of CAT sequences. Thus, the main purpose of this communication is to alert researchers working in gene regulation to be cautious when interpreting results based on heterologous reporter genes.
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http://dx.doi.org/10.1007/s00436-007-0640-6 | DOI Listing |
Intracellular protein production in bacteria is limited by the need for lysis and costly purification. A promising alternative is to engineer the host organism for protein secretion. While the serovar Typhimurium ( .
View Article and Find Full Text PDFBioorg Chem
January 2025
Helmholtz International Lab for Anti-Infectives, State Key Laboratory of Microbial Technology, Shandong University, Qingdao, Shandong 266237, China. Electronic address:
Infections of multidrug-resistant pathogens including Pseudomonas aeruginosa, cause a high risk of mortality in immunocompromised patients and underscore the need for novel natural antibacterial drugs. In this study, common phytochemicals prevalent in fruits and vegetables have been demonstrated for their ability to inhibit quorum sensing (QS) in Pseudomonas aeruginosa PAO1 (PA). Ten compounds were screened virtually by molecular docking, among which, daidzein dimethyl ether originally from Albizzia lebbeck showed the most significant inhibitory effect on the formation of biofilm and the accumulation of virulence factors, including elastase, pyocyanin and rhamnolipid in PA.
View Article and Find Full Text PDFMicroorganisms
December 2024
Department of Chemical, Biological and Environmental Engineering, Universitat Autònoma de Barcelona, Carrer de les Sitges, s/n, 08193 Bellaterra, Catalonia, Spain.
The introduction of heterologous pathways into microbial cell compartments offers several potential advantages, including increasing enzyme concentrations and reducing competition with native pathways, making this approach attractive for producing complex metabolites like fatty acids and fatty alcohols. However, measuring subcellular concentrations of these metabolites remains technically challenging. Here, we explored 3-hydroxypropionic acid (3-HP), readily quantifiable and sharing the same precursors-acetyl-CoA, NADPH, and ATP-with the above-mentioned products, as a reporter metabolite for peroxisomal engineering in the yeast .
View Article and Find Full Text PDFJ Integr Plant Biol
January 2025
Integrative Science Center of Germplasm Creation in Western China (Chongqing) Science City & Southwest University, College of Agronomy and Biotechnology, Southwest University, Chongqing, 400715, China.
The development of rapeseed with high resistance against the pathogen Sclerotinia sclerotiorum is impeded by the lack of effective resistance resources within host species. Unraveling the molecular basis of nonhost resistance (NHR) holds substantial value for resistance improvement in crops. In the present study, small RNA sequencing and transcriptome sequencing were carried out between rice (a nonhost species of S.
View Article and Find Full Text PDFInt J Biol Macromol
January 2025
College of Forestry/State Key L aboratory of North China Crop Improvement and Regulation, Hebei Agricultural University, Baoding 071000, China. Electronic address:
WD40 superfamily genes are integral to various aspects of plant growth and development. Despite the economic importance and agricultural significance of the kiwifruit (Actinidia chinensis), a comprehensive characterization of the WD40 superfamily in this species remains elusive. In this study, we identified 280 WD40-encoding genes within the kiwifruit genome and systematically analyzed their phylogenetic relationships, gene structures, functional domains, and synteny.
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