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DNA-seq of ELMSeq reporter cassette

ABSTRACT: A ELMSeq reporter cassette was created to monitor Dam levels by methylation, and introduced in the genome. Some regions upstream dam were randomized to see their effect on gene expression. So the cassette consists in: 4xGATC sites - (promoter) - random region - dam. The amplicon was spanning the whole cassette, including some bases inside the dam. The cassette was introduced in Mycoplasma pneumoniae.

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Mycoplasma pneumoniae

SUBMITTER: Jae-Seong Yang

PROVIDER: E-MTAB-5365 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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A reporter system coupled with high-throughput sequencing unveils key bacterial transcription and translation determinants.

Yus Eva E Yang Jae-Seong JS Sogues Adrià A Serrano Luis L

Nature communications 20170828 1

Quantitative analysis of the sequence determinants of transcription and translation regulation is relevant for systems and synthetic biology. To identify these determinants, researchers have developed different methods of screening random libraries using fluorescent reporters or antibiotic resistance genes. Here, we have implemented a generic approach called ELM-seq (expression level monitoring by DNA methylation) that overcomes the technical limitations of such classic reporters. ELM-seq uses D ...[more]

PMID: 28848232

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DNA-seq of an ELMSeq reporter cassette to study impact of C-terminal sequence variations on protein abundance

Project description:A ELMSeq reporter cassette was created to monitor Dam levels by methylation, and introduced in the genome. The regions of 6 nt upstream and 6 nt downstream the stop codon were randomized to study their effect on gene expression. The ELMSeq reporter cassette was composed of: promoter - dam - random N6 - stop codon TAA - random N6 - spacer - 4xGATC. The amplicon was spanning the C-terminal region. The cassette was introduced in Mycoplasma pneumoniae.

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