EC Number   |
|---|
   4.6.1.22 | chromosomal DNA from the RNase J1 conditional mutant strain used to transform Bacillus subtilis host BG1 to erythromycin resistance. The RNase J1 conditional strain also contains plasmid pMAP65, which carries extra copies of the lacI gene. Preparation and transformation of Bacillus subtilis competent cell cultures. RNase J1 transcription under control of an IPTG-inducible promoter in the RNase J1 conditional mutant strain |
| 4.6.1.22 | cloning of rnjB gene, encoding RNase J2, and/or rnjA gene expressing RNase J1 in the pET28a vector together or alone. Overexpression of C-terminal His-tagged RNase J1 and/or C-terminal His-tagged RNase J2 simultaneously or alone in Escherichia coli strain BL21 CodonPlus |
| 4.6.1.22 | gene rnjA, DNA and amino acid sequence determination and analysis, sequence comparison to Bacillus subtilis enzyme, phylogenetic analysis |
| 4.6.1.22 | gene rnjB is transcribed constitutively from a sigma A promoter. Gene rnjA is transcribed as a bicistronic transcript from a single promoter, optimal expression of RNase J1 from gene rnjA requires cotranscription and cotranslation with the upstream ykzG gene, sequence and regulatory elements upstream of the rnjA open reading frame, overview. In the absence of coupled translation, RNase J1 expression is decreased more than 5fold, transcription of the ykzG operon initiates at a sigma A promoter with a noncanonical35 box that is required for optimal transcription. In Bacillus subtilis strain SSB356, the rnjA gene is under the control of the xylose-inducible Pxyl promoter |
| 4.6.1.22 | gene rnjB, DNA and amino acid sequence determination and analysis, sequence comparison to Bacillus subtilis enzyme, phylogenetic analysis |
| 4.6.1.22 | gene rny, encoding RNase Y under control of the inducible Pspac promoter, DNA and amino acid sequence determination and analysis |
| 4.6.1.22 | into the pET28b vector for expression in Escherichia coli BL21DE3pLysS cells |
| 4.6.1.22 | overexpression of wild-type and mutant protein P proteins in Escherichia coli strain BL21 (DE3) pLysS |
| 4.6.1.22 | the wild-type ykqC gene is cloned into the pET28 vector for expression in Escherichia coli BL21 CodonPlus cells, the plasmids pMUTIN-4M, pBS-Spc and pSWEET are used for the construction of different Bacillus subtilis strains |
