Difference between revisions of "Part:BBa K4244059"

Latest revision as of 12:42, 12 October 2022

Constitutive expression of sfGFP with PelB secretion tag

sfGFP BBa_K4244004 was fused to PelB BBa_J32015 (fused at the N-terminus) to secrete sfGFP and use it as an extracellular reporter. It is under an constitutive medium strength promoter BBa_K4244057.

Secretion assay

Figure 1: Time series measurement of total, pellet and supernatant fraction of sfGFP under the PelB signal peptide expressed with constitutive promoter. Measurement was performed every two hours for eight hours. No significant fluorescence was detected in the supernatant fraction.

Discussion

No secretion was observed in Escherichia coli Nissle 1917. This may be due to the too high expression level of sfGFP under BBa_K4244057. This experiment still need adjustment and fine tuning to prove the secretion of sfGFP.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 285
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4244059 short</partinfo>
-sfGFP <partinfo>BBa_K4244004</partinfo> was fused to PelB <partinfo>BBa_J32015</partinfo> to secrete sfGFP and use it as an extracellular reporter. It is under an constitutive medium strength promoter <partinfo>BBa_K4244057</partinfo>.
+sfGFP <partinfo>BBa_K4244004</partinfo> was fused to PelB <partinfo>BBa_J32015</partinfo> (fused at the N-terminus) to secrete sfGFP and use it as an extracellular reporter. It is under an constitutive medium strength promoter <partinfo>BBa_K4244057</partinfo>.
 ====Secretion assay====
@@ Line 10: / Line 10: @@
 ====Discussion====
-No secretion was observed in ''E. coli'' Nissle. This may be due to the too high expression level of sfGFP under <partinfo>BBa_K4244057</partinfo>. This experiment still need adjustment and fine tuning to prove the secretion of sfGFP.
+No secretion was observed in ''Escherichia coli'' Nissle 1917. This may be due to the too high expression level of sfGFP under <partinfo>BBa_K4244057</partinfo>. This experiment still need adjustment and fine tuning to prove the secretion of sfGFP.