User:Josh K. Michener
Josh Michener (jkm at mit dot edu)
Currently an undergraduate at MIT, working as a UROP in the Endy Lab.
MIT Class of 2006, majoring in Chemical Engineering and Biology
Combining biological parts is currently very difficult. Parts are connected in the hopes that the levels of POPS or RIPS will match. At the moment, though, we do not have enough information to predict whether two parts will function correctly. If, for instance, we connect two inverters in series, it is quite possible that switching the output of the first inverter will not be able to switch the second. Previous attempts to measure these levels have been unsuccessful. We hope to learn from this and try again.
As such, I am building a set of POPS based screening plasmids. If all goes as planned, these plasmids will allow us to measure the transfer functions of parts (inverters or other) in units which are proportional to POPS (with the same constant of proportionality for the part's input and output). We will then try to pair up parts and verify that the behavior is as expected.
Choe et al. Nucleic Acids Res. 2005 Mar 14;33(5):e49. (PubMed Central)
- Constructed a GFP-dsRED dual fluorescence expression system. Inserted pieces of a BAC in between, searching for terminators (which would prevent dsRED expression).
Maksimow et al. Cytometry. 2002 Apr 1;47(4):243-7. (Wiley)
- Created a GFP-dsRED gene fusion to measure dual fluorescence. Showed a wide scatter (presumably machine error) even under this tightly linked expression system.
Shimada et al. J Bacteriol. 2004 Nov;186(21):7112-22. (PubMed Central)
- Created a screening plasmid with dsRED under the control of a constitutive promoter (lacUV5) and GFP under the control of a variable promoter. Used dsRED as a control for variable GFP expression.