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Transposon random mutagenesis of Mycoplasma pneumoniae M129 using ProTInSeq protocol along different concentrations and using two positive selection markers (Chloramphenicol acetyltransferase and Erythromycin esterase EreA) and a negative selection marker (barnase)

ABSTRACT: Definition of different Tn4001-derived mini-transposons carrying reporters with no translation initiation codons so they can only be expressed when fused to an endogenous protein. We used Chloramphenicol acetyltransferase (Cm) as a positive selection marker while the RNAse barnase (Barn) was used as a negative one. In this case, the mini-transposon has a constitutively expressed chloramphenicol resistance gene downstream to the Barn gene. Also, Erythromycin esterase EreA (Ery) was used to evaluate the effect of oligomerization of protein-fusion resistances, as Ery is a dimeric resistance while Cm forms a tetramer. In the positive selection, for the bacteria to survive in the presence of Chloramphenicol, the transposed reporter needs to be inserted in-frame to a genome protein-coding sequence. In the negative selection, the transposed gene should be out of frame with the genome protein-coding sequence for the bacteria to survive.

INSTRUMENT(S): Illumina HiSeq 4000

ORGANISM(S): Mycoplasma pneumoniae M129

SUBMITTER:

PROVIDER: E-MTAB-10380 | biostudies-arrayexpress |

SECONDARY ACCESSION(S): ERP128628

REPOSITORIES: biostudies-arrayexpress

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