Difference between revisions of "Part:BBa K4162026"
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[[File:T--Fudan--micro--m2-BCMO-ybbO.png|400px|thumb|none|'''Figure 2. '''Fluorescent imaging confirm the expression of both BCMO and ybbO. We observed fluorescence images of E. coli (1. Bacteria expressing BCMO-linker-StayGold ; 2. Bacteria expressing BCMO and ybbO-tdMCP-GFP ) through an Olympus fluorescence microscope with a 1.45 NA 150× oil objective. GFP and StayGold were excited by a 488 nm laser. Images were taken by the same settings for comparison.]] | [[File:T--Fudan--micro--m2-BCMO-ybbO.png|400px|thumb|none|'''Figure 2. '''Fluorescent imaging confirm the expression of both BCMO and ybbO. We observed fluorescence images of E. coli (1. Bacteria expressing BCMO-linker-StayGold ; 2. Bacteria expressing BCMO and ybbO-tdMCP-GFP ) through an Olympus fluorescence microscope with a 1.45 NA 150× oil objective. GFP and StayGold were excited by a 488 nm laser. Images were taken by the same settings for comparison.]] | ||
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| + | [[File:T--Fudan--micro--m3-6a.png|400px|thumb|none|'''Figure 3. Fluorescent imaging revealed that TEARS cluster ybbO in bacteria.'''We observed fluorescence images of E. coli (Bacteria expressing CAG-MS2, BCMO, ybbO-tdMCP-GFP, crtYEBI) through an Olympus fluorescence microscope. Four different views are shown. Please note for GFP is not required for TEARS to work. During retinol production, we used the bacteria expressing CAG-MS2, BCMO, ybbO-tdMCP-GFP, crtYEBI.]] | ||
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Latest revision as of 02:30, 14 October 2022
ribozyme + T7_RBS + BCMO + ribozyme + T7_RBS + ybbO-tdMCP
Introduction
BCMO and gene ybbO tagged with tdMCP separated by hammerhead ribozyme.
Characterization
HPLC
Figure 1. HPLC. Acetone extracted samples were subjected to HPLC. The extract from bacteria DH5α expressing CAG-MS2, BCMO, ybbO, crtYEBI (the panel above, short name 3-7) compared with the extract from bacteria expressing CAG-MS2, BCMO, ybbO-tdMCP, crtYEBI (the panel below, short name 3td7), there are two peaks in the figure between 2-4 minutes (where our standards appear as well others, detected at 325 nm), the absorbance value of the two peaks of 3td7 in this period is significantly higher than that of 3-7, which depicted the efficiency of TEARS in improving the production of retinol.
Fluorescence microscope image
Figure 2. Fluorescent imaging confirm the expression of both BCMO and ybbO. We observed fluorescence images of E. coli (1. Bacteria expressing BCMO-linker-StayGold ; 2. Bacteria expressing BCMO and ybbO-tdMCP-GFP ) through an Olympus fluorescence microscope with a 1.45 NA 150× oil objective. GFP and StayGold were excited by a 488 nm laser. Images were taken by the same settings for comparison.
Figure 3. Fluorescent imaging revealed that TEARS cluster ybbO in bacteria.We observed fluorescence images of E. coli (Bacteria expressing CAG-MS2, BCMO, ybbO-tdMCP-GFP, crtYEBI) through an Olympus fluorescence microscope. Four different views are shown. Please note for GFP is not required for TEARS to work. During retinol production, we used the bacteria expressing CAG-MS2, BCMO, ybbO-tdMCP-GFP, crtYEBI.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1018
Illegal AgeI site found at 1611 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 2101
Illegal BsaI site found at 2455