Part:BBa_K4737000
prpsM
prpsM is the promoter of 30S ribosomal subunit protein S13 of Salmonella enterica subsp. enterica serovar Typhimurium str. LT2
Usage and Biology
We used prpsM to start the expression of Lpp-OmpA-scFv, SopE-FLAG-GOx, TetR, T7 RNA polymerase, HlyA and so on.
Figure 1: The Lpp-OmpA-scFv expression pathway.
Figure 2: The SopE-FLAG-GOx expression pathway.
Figure 3: The HlyA expression pathway.
Figure 4: The T7 RNA Polymerase expression pathway.
Recombinant S. typhimurium (VNP-SopE-GOx) was grown at appropriate conditions. The proteins from the culture supernatant and bacterial lysates were prepared for an immunoblotting assay. The results of Western blot showed that VNP20009 electroporated with plasmid pFPV25.1-SopE-FLAG-GOx expressing SopE-FLAG-GOx protein successfully (Figure 5).
Figure 5: Western blot analysis shows the expression of gene of interest(SopE-FLAG-GOx) in VNP-SopE-GOx. The theoretical molecular weight of SopE-FLAG-GOx is about 76 kDa, and the theoretical molecular weight of GAPDH is about 36 kDa.
To verify the engineered bacteria can achieve the expression of HlyA, bacterial lysates were prepared from S. typhimurium strain VNP20009 carrying appropriate expression vectors for shRNA-SLC7A11 and subjected to immunoblotting analysis.HlyA expression detected by Western blotting indicates that our plasmids were successfully transferred into VNP20009 and effectively expressed (Figure 6).
Figure 6: Western blot analysis of Listeriolysin O. a.Listeriolysin O is a pore-forming protein (59kDa) encoded by gene HlyA b.GAPGH is a reference protein in cells with a molecular weight of 36 kDa.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 1
Illegal PstI site found at 34 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 34
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 1
Illegal PstI site found at 34 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 1
Illegal PstI site found at 34 - 1000COMPATIBLE WITH RFC[1000]
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