Difference between revisions of "Part:BBa K2328044"
 
Line 16: Line 16:
 
<!-- -->
 
<!-- -->
 
===Usage===
 
===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. HO-1 III is a codon-optimized version for higher expression in Escherichia coli. Besides, RBS III is used to construct the co-expression structure of HO-1 and any protein linked before RBS.
+
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. HO-1 III is a codon-optimized version for higher expression in Bifidobacterium longum. Besides, RBS III is used to construct the co-expression structure of HO-1 and any protein linked before RBS.
  
 
===Biology===
 
===Biology===

Latest revision as of 15:52, 26 October 2017


Histag.a + RBS III + HO-1 III

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 308
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 308
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 308
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 308
  • 1000
    COMPATIBLE 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. HO-1 III is a codon-optimized version for higher expression in Bifidobacterium longum. Besides, RBS III is used to construct the co-expression structure of HO-1 and any protein linked before RBS.

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.