Mycoplasma anatis, a member of the class Mollicutes, is the causative agent of a contagious infectious disease of domestic ducklings, wild birds, and eggs. Increasing reports show that coinfection of M. anatis with Escherichia coli results in substantial economic impacts on the duck farms in China. Here, we announce the first genome sequence of M. anatis. ...[more]
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. Overall design: Transcriptome profiling of 1) wild type Mycoplasma gallisepticum cell culture and mutants with transposone insertion to 5’ UTR of hypothetical protein GCW_03380, RBS of lactate dehydrogenase GCW_00390, helicase SNF2 GCW_03935, 1-deoxy-D-xylulose 5-phosphate reductoisomerase GCW_00495 or potential sigma factor GCW_00440 in exponential growth phase; 2) Mycoplasma gallisepticum cell culture under treatment with sublethal concentrations of carbonyl cyanide m-chlorophenylhydrazone (CCCP), novobiocin or tetracycline; and 3) Mycoplasma gallisepticum cell culture under heat stress at 46C during 15 min. All experiments were carried out in 2 biological replicates.
Project description:Various anti-mycoplasma drugs have different effects on cells growth We used microarrays to detail the global programme of gene expression underlying gastric cancer cells treated with anti-mycoplasma drugs and identified distinct classes of up-regulated genes during this process. Overall design: Gastric cancer cells treated with anti-mycoplasma drugs were selected for RNA extraction and hybridization on Affymetrix microarrays. We sought to obtain a disparate impact of gastric cancer cells treated with anti-mycoplasma drugs. To that end, we selected two cell lines of gastric cancer cell, AGS and BGC823, which were treated with four anti-mycoplasma drugs (A2PP, CIP, MYCO1 and MYCO2) separately.
Project description:Mycoplasmas are simple, but successful parasites that have the smallest genome of any free-living cell and are thought to have a highly streamlined cellular metabolism. Here we have undertaken a detailed metabolomic analysis of two species, Mycoplasma bovis and Mycoplasma gallisepticum, which cause economically important diseases in cattle and poultry, respectively. Untargeted GC/MS and LC/MS analyses of mycoplasma metabolite extracts revealed significant differences in the steady state levels of many metabolites in central carbon metabolism, while 13C stable isotope labelling studies revealed marked differences in carbon source utilization. These data were mapped onto in silico metabolic networks predicted from genome wide annotations. The analyses elucidated distinct differences, including a clear difference in glucose utilisation, with a marked decrease in glucose uptake and glycolysis in M. bovis compared to M. gallisepticum, which may reflect differing host nutrient availabilities. The 13C-labeling patterns also revealed several functional metabolic pathways that were previously unannotated in these species, allowing us to assign putative enzyme functions to the products of a number of genes of unknown function, especially in M. bovis. This study demonstrates the considerable potential of metabolomic analyses to assist in characterising significant differences in the metabolism of different bacterial species, and in improving genome annotation.