Difference between revisions of "Part:BBa K2253000"
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| − | The P8 constitutive promoter is natively found <i>Lactococcus lactis</i>. Although the transcriptional efficiency of this promoter has been characterized and tested in <i>Lactococcus lactis</i> and other Gram-positive bacteria, its functionality in Gram-negative species such as <i>E. coli</i> has not been recorded in the literature. We have confirmed that <i>E. coli</i> transformed with a cassette plasmid containing the P8 promoter and E2-Crimson reporter is able to successfully express the E2-Crimson red fluorescent protein. As a PhytoBrick compatible part, P8 promoter can be assembled into a transcriptional unit via Golden Gate assembly method to upregulate expression of a gene of interest in <i>E. coli</i> as well as <i>L. lactis</i>. of a gene of interest in <i>E. coli</i> as well as <i>L. lactis</i>. | + | The P8 constitutive promoter is natively found <i>Lactococcus lactis</i>. Although the transcriptional efficiency of this promoter has been characterized and tested in <i>Lactococcus lactis</i> and other Gram-positive bacteria, its functionality in Gram-negative species such as <i>E. coli</i> has not been recorded in the literature. We have confirmed that <i>E. coli</i> transformed with a cassette plasmid containing the P8 promoter and E2-Crimson reporter is able to successfully express the E2-Crimson red fluorescent protein. As a PhytoBrick compatible part, P8 promoter can be assembled into a transcriptional unit via Golden Gate assembly method to upregulate expression of a gene of interest in <i>E. coli</i> as well as <i>L. lactis</i>. of a gene of interest in <i>E. coli</i> as well as <i>L. lactis</i>. The P32 promoter and RBS sequence are natively found in the genome of <i>Lactococcus lactis</i>. However, we ordered the promoter and RBS sequence as a gBlock using the sequence information provided by Zhu et al. (2015). |
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Revision as of 01:27, 25 October 2017
Constitutive P8 promoter and RBS composite
The P8 constitutive promoter is natively found Lactococcus lactis. Although the transcriptional efficiency of this promoter has been characterized and tested in Lactococcus lactis and other Gram-positive bacteria, its functionality in Gram-negative species such as E. coli has not been recorded in the literature. We have confirmed that E. coli transformed with a cassette plasmid containing the P8 promoter and E2-Crimson reporter is able to successfully express the E2-Crimson red fluorescent protein. As a PhytoBrick compatible part, P8 promoter can be assembled into a transcriptional unit via Golden Gate assembly method to upregulate expression of a gene of interest in E. coli as well as L. lactis. of a gene of interest in E. coli as well as L. lactis. The P32 promoter and RBS sequence are natively found in the genome of Lactococcus lactis. However, we ordered the promoter and RBS sequence as a gBlock using the sequence information provided by Zhu et al. (2015).
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]
References
- Zhu, D. et al. Isolation of strong constitutive promoters from Lactococcus lactis subsp. Lactis N8. FEMS Microbiol Lett. 363(16): pii: fnv107 (2015).