Difference between revisions of "Part:BBa K3606812"

Revision as of 02:33, 5 December 2020

mcbG

Usage and Biology:

McbG is responsible for immunity against 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.

Results:

We've constructed the plasmid for mcbB and mcbD expression, yet the results of colony PCR is false positive thus further experiment for the fundamental validation of the antimicrobial peptide(mccb17) expression is needed. Figure1. gel map for mcbA, mcbC, mcbE and mcbF

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]

@@ Line 10: / Line 10: @@
 <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
+We've constructed the plasmid for mcbB and mcbD expression, yet the results of colony PCR is false positive thus further experiment for the fundamental validation of the antimicrobial peptide(mccb17) expression is needed.
+https://2020.igem.org/wiki/images/f/f3/T--Fudan--Poster_Mcb-gels.jpg
-Comparing the 5th and 6th lanes, after IPTG induction, there is a clear band, which is the product of McbG expression.
+Figure1. gel map for mcbA, mcbC, mcbE and mcbF
 <!-- Add more about the biology of this part here