Project description:We have engineered synthetic gene switches to control and limit Mycoplasma growth for biosafety containment applications. Mycoplasmas have high mutation rates and, the accumulation of mutations that inactivate the circuit is expected. However, the question is how resilient is the kill-switch to mutation and whether it is more sensitive to the accumulation of mutations. Therefore, we did the whole-genome sequencing of the three Mycoplasma biosafety strains, designed in our study, at different passages (p2, p3 and p15) or after IPTG-treatment at passage 3 (p3IPTG)
Project description:Mycoplasma gallisepticum is a convenient model object for studying the regulation of transcription because it has a reduced genome, lack of cell wall and many metabolic pathways, and also easy to culture and non-pathogenic to humans. For rapid investigation of gene expression we developed microarray design including 3 366 probes for 678 genes. They included 665 protein coding sequences and 13 antisense RNAs from 816 genes and 17 ncRNAs present in Mycoplasma gallisepticum. This work was carried out transcriptomic profiling for different types of effects on the expression of genes of Mycoplasma gallisepticum: 1) genetic knock-out mutants; 2) cell culture exposed to sublethal concentrations of antibiotics; and 3) well-characterized heat stress effect. The study was performed on Agilent one-color microarray with custom design and random-T7 polymerase primer for cDNA synthesis. Using set of different probes for each gene or ncRNA allows to increase accuracy of gene expression quality.
Project description:Overall design Hi-C experiments were performed on untreated wild type cells at stationary and drug-treated cells (Novobiocin) of Mycoplasma Pneumoniae MPN129. We studied the chromosome organization of the genome-reduced bacterium, Mycoplasma pneumoniae, which has minimal genetic components and lacks several structural DNA-binding proteins. Platforms : Illumina HiSeq 2000 (Mycoplasma Pneumoniae MPN129)
Project description: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.
