Difference between revisions of "Part:BBa K3606811"

Revision as of 02:02, 28 October 2020

mcbF

McbF works as channel protein responsible for efflux of MccB17.

Design:

Since we encountered difficulties in the expression of McbABCDEFG at the beginning, we decided to check whether the McbA/B/C/D/E/F/G genes can be expressed and translated separately. We successfully cloned McbG into the pGEX plasmid and transcribed it into BL21. After 5 hours of induction of McbG expression with IPTG, the whole protein SDS-PAGE was used to detect whether the McbG product was produced.

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
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 Since we encountered difficulties in the expression of McbABCDEFG at the beginning, we decided to check whether the McbA/B/C/D/E/F/G genes can be expressed and translated separately. We successfully cloned McbG into the pGEX plasmid and transcribed it into BL21. After 5 hours of induction of McbG expression with IPTG, the whole protein SDS-PAGE was used to detect whether the McbG product was produced.
-<h2>Results:</h2>
-https://2020.igem.org/wiki/images/2/22/T--Fudan--img_McbA-E-G-BCD.jpeg
-Figure 1. SDS-PAGE result. lane 1: mcbA  lane 2: mcbA+IPTG lane 3: mcbE  lane 4: mcbE+IPTG  lane 5: mcbG  lane 6: mcbG+IPTG  lane 7: mcbBCD  lane 8: mcbBCD+IPTG lane 9: pGEX+ IPTG
-Comparing the 5th and 6th lanes, after IPTG induction, there is a clear band, which is the product of McbG expression.