Difference between revisions of "Part:BBa K4015003"
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Gene III encodes pIII, which is synthesized from 18 amino acids. It is an amino-terminal signal sequence that necessitates insertion of the Sec system into the membrane. The signal sequence is removed after passage through the inner membrane while most of the protein will be retained in the periplasmic space. | Gene III encodes pIII, which is synthesized from 18 amino acids. It is an amino-terminal signal sequence that necessitates insertion of the Sec system into the membrane. The signal sequence is removed after passage through the inner membrane while most of the protein will be retained in the periplasmic space. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | + | In our project, we pick three signal peptides to help secrete our target proteins, which are HlyA、GeneIII and DsbA . | |
| + | We used p47 vector containing GFP to compare the effectiveness of these signal peptides. p47-GFP-HlyA displayed a lower level of fluorescence comparing to p47-GeneIII-GFP, suggesting HlyA is more effective in secretion since more GFP has entered the medium. p47-DsbA-GFP showed no fluorescence, meaning protein expression was inhibited or the folding of the protein has failed. | ||
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| + | <div>Citation:<div> | ||
| + | 1. Linton, E., Walsh, M. K., Sims, R. C., & Miller, C. D. (2011). Translocation of green fluorescent protein by comparative analysis with multiple signal peptides. Biotechnology Journal, 7(5), 667-676. https://doi.org/10.1002/biot.201100158 | ||
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| + | 2. Boeke, J. D., Model, P., & Zinder, N. D. (1982). Effects of bacteriophage f1 gene III protein on the host cell membrane. Molecular & general genetics : MGG, 186(2), 185–192. https://doi.org/10.1007/BF00331849 | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K4015003 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4015003 SequenceAndFeatures</partinfo> | ||
Latest revision as of 13:43, 20 October 2021
Gene III
Gene III encodes pIII, which is synthesized from 18 amino acids. It is an amino-terminal signal sequence that necessitates insertion of the Sec system into the membrane. The signal sequence is removed after passage through the inner membrane while most of the protein will be retained in the periplasmic space.
Usage and Biology
In our project, we pick three signal peptides to help secrete our target proteins, which are HlyA、GeneIII and DsbA . We used p47 vector containing GFP to compare the effectiveness of these signal peptides. p47-GFP-HlyA displayed a lower level of fluorescence comparing to p47-GeneIII-GFP, suggesting HlyA is more effective in secretion since more GFP has entered the medium. p47-DsbA-GFP showed no fluorescence, meaning protein expression was inhibited or the folding of the protein has failed.
1. Linton, E., Walsh, M. K., Sims, R. C., & Miller, C. D. (2011). Translocation of green fluorescent protein by comparative analysis with multiple signal peptides. Biotechnology Journal, 7(5), 667-676. https://doi.org/10.1002/biot.201100158
2. Boeke, J. D., Model, P., & Zinder, N. D. (1982). Effects of bacteriophage f1 gene III protein on the host cell membrane. Molecular & general genetics : MGG, 186(2), 185–192. https://doi.org/10.1007/BF00331849
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]