@@ -35,6 +35,7 @@ HIPs are regulated by the Fumarate and Nitrate Reduction regulator (FNR)[$^1$].
...
@@ -35,6 +35,7 @@ HIPs are regulated by the Fumarate and Nitrate Reduction regulator (FNR)[$^1$].
Thus, on the base of wild type PepT promoter, we strived to change the intensity of its expression by mutating TATA box, mutating FNR box, and changing the RBS. We use the PJUMP41-2A (sfGFP) plasmid (BBa_J428365, this plasmid referred below are abbreviated as O12, as it corresponds to the O12 well on the biobrick) as our vector This plasmid is characterized as a low-copy plasmid with spectinomycin resistance and carries the BBa_J23100 promoter along with the sfGFP sequence. Through reverse PCR and gibson assembly (details are shown in “Engineering”), we successfully replaced the J23100 promoter and RBS with the following 10 promoters + RBS (see **BBa_K4713116 - BBa_K4713125**):
Thus, on the base of wild type PepT promoter, we strived to change the intensity of its expression by mutating TATA box, mutating FNR box, and changing the RBS. We use the PJUMP41-2A (sfGFP) plasmid (BBa_J428365, this plasmid referred below are abbreviated as O12, as it corresponds to the O12 well on the biobrick) as our vector This plasmid is characterized as a low-copy plasmid with spectinomycin resistance and carries the BBa_J23100 promoter along with the sfGFP sequence. Through reverse PCR and gibson assembly (details are shown in “Engineering”), we successfully replaced the J23100 promoter and RBS with the following 10 promoters + RBS (see **BBa_K4713116 - BBa_K4713125**):
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**Table 1 | The promoter + RBS sequences used in screening.**
**Table 1 | The promoter + RBS sequences used in screening.**
@@ -363,4 +364,4 @@ This design can be utilized to optimize therapeutic bacteria administration to m
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@@ -363,4 +364,4 @@ This design can be utilized to optimize therapeutic bacteria administration to m
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