Difference between revisions of "Part:BBa K4162026"
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BCMO and gene ''ybbO'' tagged with tdMCP separated by hammerhead ribozyme.
 
BCMO and gene ''ybbO'' tagged with tdMCP separated by hammerhead ribozyme.
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===Characterization===
  
 
====HPLC====
 
====HPLC====
  
 
[[File:T--Fudan--026--027.png|400px|thumb|none|'''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.]]
 
[[File:T--Fudan--026--027.png|400px|thumb|none|'''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.]]
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====Fluorescence microscope image====
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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.]]
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 08:12, 13 October 2022


ribozyme + T7_RBS + BCMO + ribozyme + T7_RBS + ybbO-tdMCP

Introduction

2022 Fudan

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.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1018
    Illegal AgeI site found at 1611
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2101
    Illegal BsaI site found at 2455