DNA sequencing to validate random deletions in Mycoplasma pneumoniae
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ABSTRACT: We present LoxTnSeq, a new methodology to generate and catalogue libraries of genome reduction mutants. LoxTnSeq combines random integration of Lox sites by transposon mutagenesis, and the generation of mutants via cre recombinase, catalogued via deep-sequencing. When LoxTnSeq was applied to the naturally genome reduced bacterium Mycoplasma pneumoniae, we obtained a mutant pool containing 285 unique deletions. These deletions spanned from >50 bp to 28 Kb, which represent 21% of the total genome. LoxTnSeq also highlighted large regions of non-essential genes that could be removed simultaneously, and other similar regions that could not, providing a guide for future genome reductions.
INSTRUMENT(S): Illumina HiSeq 2500
ORGANISM(S): Mycoplasma pneumoniae
SUBMITTER: Maria Lluch-Senar
PROVIDER: E-MTAB-9582 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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