ARF-TSS: an alternative method for identification of transcription start site in bacteria

Current methods for identifying transcription start sites (TSSs) of specific genes in bacteria usually require adaptors or radioactive labeling. These approaches can be technically demanding and environmentally unfriendly. Here we present a method for identifying TSS called ARF-TSS, which is based o...

وصف كامل

التفاصيل البيبلوغرافية
الحاوية / القاعدة:BioTechniques
المؤلفون الرئيسيون: Chao Wang, Jasmine Lee, Yinyue Deng, Fei Tao, Lian-Hui Zhang
التنسيق: مقال
اللغة:الإنجليزية
منشور في: Taylor & Francis Group 2012-04-01
الموضوعات:
transcripts
promoter
reverse transcription
transcriptional initiation
mRNA
Pseudomonas aeruginosa
الوصول للمادة أونلاين:https://www.future-science.com/doi/10.2144/000113858
الوصف
الملخص:Current methods for identifying transcription start sites (TSSs) of specific genes in bacteria usually require adaptors or radioactive labeling. These approaches can be technically demanding and environmentally unfriendly. Here we present a method for identifying TSS called ARF-TSS, which is based on cDNA generation, circularization, PCR amplification, and DNA sequencing to determine the 5′-end of transcripts, thus circumventing the need for adaptors and radioactive labeling. We validated the method using the gene lasI from the bacterial pathogen Pseudomonas aeruginosa. Our results show that ARF-TSS could be a good alternative to traditional methods for bacterial TSS analysis.
تدمد:0736-6205
1940-9818

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