Difference between revisions of "Part:BBa K769001"

Revision as of 15:52, 14 October 2019

PompC-RBS-GFP-Double terminator

When OmpC promotor is activated by phosphorylated OmpR, GFP is expressed.

Usage and Biology

Characterization by team IGEM19_UI_Indonesia

Highest expression of GFP in TOP10 E. coli strain

Expression of GFP in TOP10 showed a sharp increase at 2.5 mg/20mL NaCl concentration compared to 2.0 mg/20mL concentration. This is contradicting our thinking that TOP10 will have a lower expression compared to BL21 because BL21 is a protein expression bacteria. A possible reason for this phenomenon is that there are differences in ompC promoter regulation in different bacterial strains. According to a characterization done by SDU-Denmark, it is found that OmpR-regulated promoter showed a high leaky expression in high copy vectors (e.g. TOP10) compared to low copy vectors (e.g. BL21) confirming our findings in our characterization. From the results, DH5a then BL21 showed a gradual increase in expression with increasing NaCl concentrations in LB medium. This indicates that DH5a and BL21 has a much more efficient control of gene expression with the ompC promoter.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 656
Illegal XhoI site found at 557
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+Characterization by team IGEM19_UI_Indonesia
+Highest expression of GFP in TOP10 E. coli strain
+Expression of GFP in TOP10 showed a sharp increase at 2.5 mg/20mL NaCl concentration compared to 2.0 mg/20mL concentration. This is contradicting our thinking that TOP10 will have a lower expression compared to BL21 because BL21 is a protein expression bacteria. A possible reason for this phenomenon is that there are differences in ompC promoter regulation in different bacterial strains. According to a characterization done by SDU-Denmark, it is found that OmpR-regulated promoter showed a high leaky expression in high copy vectors (e.g. TOP10) compared to low copy vectors (e.g. BL21) confirming our findings in our characterization. From the results, DH5a then BL21 showed a gradual increase in expression with increasing NaCl concentrations in LB medium. This indicates that DH5a and BL21 has a much more efficient control of gene expression with the ompC promoter.
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