Part:BBa_K2328003
HO-1 II codon-optimized for intestinal bacteria
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]
Usage
In order to fluoresce, smURFP must be combined with biliverdin (BV). HO-1 is the gene of the precursor of biliverdin. HO-1 can use the materials of the E.coil to produce biliverdin. So we want to construct a plasmid which can both express the smURFP gene and HO-1 gene. Through this construction, we can achieve the co-expression in the E.coil. Both the smURFP and biliverdin are produced by E.coil, so they can connect directly within the E.coil to produce fluorescence under the wavelength of 642 nm without adding BV additionally. Besides, this is a codon-optimized version for higher HO-1 expression in host intestinal bacteria.
Biology
One of our methods is co-expression. Because the HO-1 needs to use oxygen to produce BV, it is adoptable in E.coil which is a kind of facultative anaerobic bacteria. And the HO-1 gene is from the Block Library. Both the smURFP and biliverdin are produced by E.coil, so they can connect directly within the E.coil and we can achieve the co-expression in the E.coil.
Reference
[1] Dong Chen , Jason D Brown , Yukie Kawasaki , Jerry Bommer and Jon Y Takemoto . Scalable production of biliverdin IXα by Escherichia coli. [J].BMC Biotechnology, 2012.
[2] Rodriguez EA, Tran GN, Gross LA, Crisp JL, Shu X, Lin JY, Tsien RY. A far-red fluorescent protein evolved from a cyanobacterial phycobiliprotein. Nat Methods. 2016 Sep;13(9):763-9.
[3] heme oxygenase (ho1) from Synechocystis Part:BBa_I15008
[4] HO-1 I (without a barcode) Part:BBa_K2328062
[5] HO-1 III codon-optimized for Bifidobacterium longum Part:BBa_K2328004
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