Part:BBa_K3606814
mcbBCD
This part is an antibiotic coding gene cluster with proteins for MccB17 maturation.
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:
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.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 2062
Illegal PstI site found at 2095 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 2062
Illegal PstI site found at 2095 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1969
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 2062
Illegal PstI site found at 2095 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 2062
Illegal PstI site found at 2095
Illegal NgoMIV site found at 1724
Illegal AgeI site found at 1896 - 1000COMPATIBLE WITH RFC[1000]
proteins |