Difference between revisions of "Team:BJRS China/Improve"
m |
|||
| Line 91: | Line 91: | ||
<ul class="dropdown-menu"> | <ul class="dropdown-menu"> | ||
<li><a href="#">MEMBERS</a></li> | <li><a href="#">MEMBERS</a></li> | ||
| − | <li><a href="https://2018.igem.org/Team:BJRS_China/ | + | <li><a href="https://2018.igem.org/Team:BJRS_China/Notebooks">NOTEBOOKS</a></li> |
<li><a href="#">PROTOCOL</a></li> | <li><a href="#">PROTOCOL</a></li> | ||
<li><a href="#">ACKNOWLEDGEMENT</a></li> | <li><a href="#">ACKNOWLEDGEMENT</a></li> | ||
Revision as of 07:03, 15 October 2018
Improvement
Design
Our project used surface display system, and we choose the INP-YFP expressiong part BBa_K523013 built by Edinburgh 2011 to construct the INP-mediated surface display for our interest protein. While testing the surface display efficiency of this part, we found that the fluorescence intensity was too weak both under the uv-light（adjusted to the same OD600）(Fig.2, middle) and under the super resolution microscope(Fig.3, middle).
To characterize the surface display efficiency of INP better, we added a strong promoter BBa_J23104 and RBS BBa_B0032 to the upstream of INP, and changed the YFP to sfGFP, to construct our parts BBa_K2833009(Fig.1)
Results
The overnight cultured bacteria expressing GFP, INP-YFP and INP-GFP respectively were firstly adjusted to OD600 = 1.0 to assure the same cell counts, then 1 mL bacteria solution of each was centrifuged to get the sediment of whole cell. From the fluorescence strength of the of whole cell(E.coli, DH5α) precipitation under UV-light we can see that the BBa_K523013 transformed cells showed the weakest fluorescence, suggesting that our new construct can give out stronger fluorescence.
