Difference between revisions of "Part:BBa K1642004"
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− | The SD-like sequence derived from the cpcB gene of Synechococcus sp. PCC7002, 5′-UAUAAGUAGGAGAUAAAAAC-3′, is to be introduced at upstream of the start codon of the genes under the PcpcG2. | + | The SD-like sequence derived from the cpcB gene of ''Synechococcus sp.'' PCC7002, 5′-UAUAAGUAGGAGAUAAAAAC-3′(Abe, K. et al. 2014), is to be introduced at upstream of the start codon of the genes under the PcpcG2. |
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+ | Ribosome binding sites(RBS) control protein expression level to a great extent. The PcpcG2 promoter does not have a typical Shine-Dalgarno (SD) sequence upstream of the start codon. Reported weak expression may be due to the lack of a SD-like sequence, so we add an SD sequence on the gene under PcpcG2, hoping that the gene will express on a relatively high level. | ||
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+ | Highly expressed photosynthesis-related proteins in cyanobacteria, the cpcB encoded c-phycocyanin, has in their gene’s 5′-UTR the SD-like sequences AGGAG respectively. The SD-like sequence from cpcB is completely complementary to the 3′ region of 16S rRNA of ''Synechocystis sp.'' PCC6803 and is highly conserved among various cyanobacteria. | ||
+ | |||
+ | Reported about 15-fold higher GFPuv-derived fluorescence intensity resulted from the addition of the SD-like sequence to PcpcG2 proved that the SD-like sequence function strongly in ''Synechocystis sp.'' PCC6803. | ||
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Latest revision as of 00:05, 19 September 2015
SD sequence of cpcB
The SD-like sequence derived from the cpcB gene of Synechococcus sp. PCC7002, 5′-UAUAAGUAGGAGAUAAAAAC-3′(Abe, K. et al. 2014), is to be introduced at upstream of the start codon of the genes under the PcpcG2.
Ribosome binding sites(RBS) control protein expression level to a great extent. The PcpcG2 promoter does not have a typical Shine-Dalgarno (SD) sequence upstream of the start codon. Reported weak expression may be due to the lack of a SD-like sequence, so we add an SD sequence on the gene under PcpcG2, hoping that the gene will express on a relatively high level.
Highly expressed photosynthesis-related proteins in cyanobacteria, the cpcB encoded c-phycocyanin, has in their gene’s 5′-UTR the SD-like sequences AGGAG respectively. The SD-like sequence from cpcB is completely complementary to the 3′ region of 16S rRNA of Synechocystis sp. PCC6803 and is highly conserved among various cyanobacteria.
Reported about 15-fold higher GFPuv-derived fluorescence intensity resulted from the addition of the SD-like sequence to PcpcG2 proved that the SD-like sequence function strongly in Synechocystis sp. PCC6803.
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]