E-MTAB-8431
Whole track RNAseq of two different integrative vectors: pSET152::PrpsJ:acbC and pGUS::PrpsJ:gusA. in Actinoplanes sp. SE50/110.
The whole coding RNA of Actinoplanes sp. SE50/110 mutants containing two different integrative vectors (pSET152::PrpsJ:acbC and pGUS::PrpsJ:gusA) were sequenced. Both vectors are integrated via a phiC31 integrase (Bierman et al. 1992) into the genetic locus ACSP50_6589 (former: acpl_6602) (Gren et al. 2016). For expression of the gene of interest (either gusA or acbC) the native promoter of the ribosomal gene rpsJ (ACSP50_0690) was used. The pGUS and pSET152 backbone mainly differ by the integration of a spectinomycin resistance cassette aadA in the pGUS-vector. The cassette is flanked by two T4-terminators to block the transcription of the goi from upstream promoters (Myronovskyi et al. 2011). As shown by this experiment, the two T4-terminators block transcription and prevent read-through efficiently in pGUS, whereas continuous tracks can be observed in pSET152, which extent from int to acbC.
The whole coding RNA of Actinoplanes sp. SE50/110 mutants containing two different integrative vectors (pSET152::PrpsJ:acbC and pGUS::PrpsJ:gusA) were sequenced. Both vectors are integrated via a phiC31 integrase (Bierman et al. 1992) into the genetic locus ACSP50_6589 (former: acpl_6602) (Gren et al. 2016). For expression of the gene of interest (either gusA or acbC) the native promoter of the ribosomal gene rpsJ (ACSP50_0690) was used. The pGUS and pSET152 backbone mainly differ by the integration of a spectinomycin resistance cassette aadA in the pGUS-vector. The cassette is flanked by two T4-terminators to block the transcription of the goi from upstream promoters (Myronovskyi et al. 2011). As shown by this experiment, the two T4-terminators block transcription and prevent read-through efficiently in pGUS, whereas continuous tracks can be observed in pSET152, which extent from int to acbC.