Difference between revisions of "Part:BBa K4183005"
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Addition of kp-sp signal peptide to the N-terminal end of GFP allows it to be secreted extracellularly. | Addition of kp-sp signal peptide to the N-terminal end of GFP allows it to be secreted extracellularly. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
We added a signal peptide (kp-sp, BBa_K2981006) from Actinobacillus at the N-terminal end of GFP (BBa_E0040) to construct a new part (BBa_K4183005) that allows GFP to be secreted extracellularly by E. coli. | We added a signal peptide (kp-sp, BBa_K2981006) from Actinobacillus at the N-terminal end of GFP (BBa_E0040) to construct a new part (BBa_K4183005) that allows GFP to be secreted extracellularly by E. coli. | ||
We incubated E. coli containing BBa_K4183005 overnight, and centrifuged 2 ml of the bacterial solution. We subsequently observed the supernatant under UV irradiation. The results showed (Figure 1) that the supernatant of the experimental group emitted green fluorescence compared with the control group without kp-sp (BBa_K2981006), demonstrating that GFP containing kp-sp was secreted outside the cell. | We incubated E. coli containing BBa_K4183005 overnight, and centrifuged 2 ml of the bacterial solution. We subsequently observed the supernatant under UV irradiation. The results showed (Figure 1) that the supernatant of the experimental group emitted green fluorescence compared with the control group without kp-sp (BBa_K2981006), demonstrating that GFP containing kp-sp was secreted outside the cell. | ||
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[[File:HS_China_improvement-1.png|900px|thumb|none|alt=Kp-Sp amilCP.|Figure 1. Secretion of GFP without (left)/with (right) addition of signal peptide (kp-sp).]] | [[File:HS_China_improvement-1.png|900px|thumb|none|alt=Kp-Sp amilCP.|Figure 1. Secretion of GFP without (left)/with (right) addition of signal peptide (kp-sp).]] | ||
Latest revision as of 14:34, 11 October 2022
Addition of kp-sp signal peptide at the N-terminal end of GFP
Addition of kp-sp signal peptide to the N-terminal end of GFP allows it to be secreted extracellularly.
Usage and Biology
We added a signal peptide (kp-sp, BBa_K2981006) from Actinobacillus at the N-terminal end of GFP (BBa_E0040) to construct a new part (BBa_K4183005) that allows GFP to be secreted extracellularly by E. coli.
We incubated E. coli containing BBa_K4183005 overnight, and centrifuged 2 ml of the bacterial solution. We subsequently observed the supernatant under UV irradiation. The results showed (Figure 1) that the supernatant of the experimental group emitted green fluorescence compared with the control group without kp-sp (BBa_K2981006), demonstrating that GFP containing kp-sp was secreted outside the cell.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 966