into the MscI site of pUI1087, generating pKS7, moving of the hemF sequences from pKS7 into pBBR1MCS2, creating pKS9, BamHI DNA fragment deleted from pKS9, omega Sp-St resistance cassette inserted at the remaining unique BamHI site, generating pKS11, from pKS11 3135-bp SmaI fragment containing delta hemF::omegaSpr-Str moved into pSUP202, replacing the vector sequences contained on a 1,348-bp ScaI fragment and creating pKS13, hemF::lacZ transcription reporter plasmid pJZ84 constructed by first inserting a 717-bp SmaI-NcoI DNA fragment from pKS9 into pUI1087, using the flanking PstI-XbaI vector restriction sites, the hemFequences are positioned and correctly oriented in front of a promoterless lacZ gene in plasmid vector pCF1010 |
Cereibacter sphaeroides |