Part:BBa_K4183006
Construct lacZ and kfiC into the same expression frame.
Construct lacZ and kfiC into the same expression frame.Promoter is Plac(R0010)
Usage and Biology
From the above experimental data, we obtained the engineered bacteria that can secrete lacZ into the extracellular and have some acid resistance. Therefore, we planned to combine the two, and thus developed and tested the first generation of engineered bacterium-lacMAN1.0 for lactos degradation.
The results showed that the lacZ secretion capacity and acid tolerance of lacMan1.0 were reduced to different degrees (Figure 1). Only 13.33% of lacZ was secreted into the extracellular compartment, lower than the previous 21.93%; its growth remained slow at pH=4 but it barely survived at pH≤3. We think that this is caused by BBa_R0010 exhibiting insufficient promoter strength for an overly long expression frame.
In addition, the induction of IPTG is extremely difficult in the intestine. Therefore, we planned to replace BBa_R0010 with the current promoter BBa_J23119, which is more strongly expressed, and deleted the original lacI element on the pET28a plasmid to reduce the replication pressure of bacteria.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 3630
Illegal PstI site found at 4762 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 3630
Illegal PstI site found at 4762 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 3630
Illegal BglII site found at 4212
Illegal BamHI site found at 4565 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 3630
Illegal PstI site found at 4762 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 3630
Illegal PstI site found at 4762 - 1000COMPATIBLE WITH RFC[1000]
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