tailieunhanh - Báo cáo y học: "Global fitness profiling of fission yeast deletion strains by barcode sequencing"

Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học Wertheim cung cấp cho các bạn kiến thức về ngành y đề tài: Global fitness profiling of fission yeast deletion strains by barcode sequencing. | Han et al. Genome Biology 2010 11 R60 http 2010 11 6 R60 w Genome Biology METHOD _ Open Access Global fitness profiling of fission yeast deletion strains by barcode sequencing Tian Xu Han Xing-Ya Xu Mei-Jun Zhang Xu Peng and Li-Lin Du Abstract A genome-wide deletion library is a powerful tool for probing gene functions and one has recently become available for the fission yeast Schizosaccharomycespombe. Here we use deep sequencing to accurately characterize the barcode sequences in the deletion library thus enabling the quantitative measurement of the fitness of fission yeast deletion strains by barcode sequencing. Background Over the past decade the availability of whole genome sequences for several major model organisms has spurred the development of many powerful reverse genetics approaches and as a consequence brought about dramatic changes to the way gene functions are analyzed. The ultimate reverse genetics tool whole-genome deletion mutant libraries were first created for the budding yeast Saccharomyces cerevisiae 1 2 . This resource allows all predicted open reading frames in the budding yeast genome to be studied by analyzing the phenotypes of their deletion mutants. Numerous screens have been conducted with the budding yeast deletion libraries to uncover new genes involved in various biological pathways 3 . In addition new approaches based on the deletion libraries such as synthetic genetic array analysis have been developed to map global genetic interaction networks 4 . The utility of the deletion libraries goes even beyond studying gene functions as profiling drugsensitive yeast mutants has allowed the targets of therapeutic compounds to be defined 5-8 . The construction of the budding yeast deletion libraries incorporated the ingenious idea of molecular barcodes which are a pair of 20-nucleotide-long unique DNA sequences flanking each deletion cassette 9 . The two barcodes for each gene are called uptag barcode upstream of the KanMX .