We describe a transient transfection protocol for cultured Leishmania major promastigotes, utilizing Escherichia coli genes encoding /?-galactosidase and B-glucuronidase inserted into an expression vector derived from the dihydrofolate reductasethymidylate synthase locus. Less than 0.1 pg of either reporter enzyme can be detected with a simple fluorimetric assay, and transfection of 10 pg of ei...

Reporter genes integrated into the genome are a powerful tool to reveal effects of regulatory elements and local chromatin context on gene expression. However, so far such reporter assays have been of low throughput. Here, we describe a multiplexing approach for the parallel monitoring of transcriptional activity of thousands of randomly integrated reporters. More than 27,000 distinct reporter ...

One of steroid hormones, ecdysteroid is a key regulatory factor, controlling the development and molting in arthropod. Molecular target of ecdysteroids is a receptor complex, composed of the ecdysone receptor (EcR) and ultraspiracle (USP). To characterize the transactivation potentials of crustacean EcR by environmental pollutants, we established an in vitro reporter gene assay using insect cel...

Serotonin (5-HT) receptors are valuable molecular targets for antipsychotic drug discovery. Current reported methods for detecting 5-HT receptors, such as cAMP accumulation and calcium influx assay, are often demanding specialized instruments and inconvenient. The luciferase reporter gene assay, based on the responsible-element-regulated expression of luciferase, has been widely applied in the ...

Bioluminescence reporter proteins have been widely used in the development of tools for monitoring biological events in living cells. In this study, we describe the development of a reporter system with secreted Cypridina noctiluca luciferase (CLuc) for a pharmacological assay that is based on targeted promoter activity. A model cell line was established with Rat-1 fibroblasts expressing CLuc d...

Industrialisation, directly or indirectly, exposes humans to various xenobiotics. The increased magnitude of chemical pesticides and toxic heavy metals in the environment, as well their intrusion into food chain, seriously threatens human health. Therefore, surveillance xenobiotics is crucial for social safety security. Online investigation by traditional methods not sufficient detection identi...

We have developed a specific and sensitive nucleic acid amplification assay that is suitable for routine gene detection. The assay is based on a novel molecular genetic strategy in which two different RNA probes are hybridized to adjacent positions on a target nucleic acid and then ligated to form an amplifiable reporter RNA. The reporter RNA is then replicated up to a hundred billion-fold in a...

We present a simple assay to examine effects of compounds on virulence gene expression in the human pathogen Staphylococcus aureus. The assay employs transcriptional reporter strains carrying lacZ fused to central virulence genes. Compounds affecting virulence gene expression and activity of the agr locus are scored based on color change in the presence of a chromogenic beta-galactosidase subst...

a stable agrobacterium-mediated transformation system was established using bialaphos as the selective agent in soybeans [glycine max (l.) merr.]. the cotyledonary node explants of the soybean cultivar ‘ny-1001’ were inoculated with the agrobacterium tumefaciens strain eha105, harboring the vector pcambia3301 containing the gus gene as the reporter gene and the bar gene conferring bialaphos res...

Many studies using reporter assays have demonstrated that 3' untranslated regions (3'-UTRs) regulate gene expression by controlling mRNA stability and translation. Due to intrinsic limitations of heterologous reporter assays, we sought to develop a gene editing approach to investigate the regulatory activity of 3'-UTRs in their native context. We initially used dual-CRISPR (clustered, regularly...