Quantifying promoter activity during the developmental cycle of Chlamydia trachomatis
| Autor: | Yan Zhang, Li Shen, Hiba Elaasar, Yanguang Cong, Leiqiong Gao, Yuqi Xian, Ziyu Hua |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Transcription Genetic 030106 microbiology Green Fluorescent Proteins Chlamydia trachomatis Biology medicine.disease_cause Article Flow cytometry Green fluorescent protein 03 medical and health sciences Rapid amplification of cDNA ends Bacterial Proteins Transcription (biology) Gene expression medicine Promoter Regions Genetic Regulation of gene expression Multidisciplinary medicine.diagnostic_test Gene Expression Regulation Developmental Promoter Gene Expression Regulation Bacterial Molecular biology 030104 developmental biology Mutation |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | Chlamydia trachomatis is an important human pathogen that undergoes a characteristic development cycle correlating with stage-specific gene expression profiles. Taking advantage of recent developments in the genetic transformation in C. trachomatis, we constructed a versatile green fluorescent protein (GFP) reporter system to study the development-dependent function of C. trachomatis promoters in an attempt to elucidate the mechanism that controls C. trachomatis adaptability. We validated the use of the GFP reporter system by visualizing the activity of an early euo gene promoter. Additionally, we uncovered a new ompA promoter, which we named P3, utilizing the GFP reporter system combined with 5′ rapid amplification of cDNA ends (RACE), in vitro transcription assays, real-time quantitative RT-PCR (RT-qPCR) and flow cytometry. Mutagenesis of the P3 region verifies that P3 is a new class of C. trachomatis σ66-dependent promoter, which requires an extended −10 TGn motif for transcription. These results corroborate complex developmentally controlled ompA expression in C. trachomatis. The exploitation of genetically labeled C. trachomatis organisms with P3-driven GFP allows for the observation of changes in ompA expression in response to developmental signals. The results of this study could be used to complement previous findings and to advance understanding of C. trachomatis genetic expression. |
| Databáze: | OpenAIRE |
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