Difference between revisions of "Part:BBa K3606807"
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McbB works as synthase responsible for the maturation of MccB17. | McbB works as synthase responsible for the maturation of MccB17. | ||
<h2>Design:</h2> | <h2>Design:</h2> | ||
| − | 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 | + | 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 McbB into the pGEX plasmid and transcribed it into BL21. After 5 hours of induction of McbB expression with IPTG, the whole protein SDS-PAGE was used to detect whether the McbA product was produced. |
<h2>Results:</h2> | <h2>Results:</h2> | ||
Revision as of 01:54, 28 October 2020
mcbB
Usage and Biology:
McbB works as synthase responsible for the maturation 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 McbB into the pGEX plasmid and transcribed it into BL21. After 5 hours of induction of McbB expression with IPTG, the whole protein SDS-PAGE was used to detect whether the McbA 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 1th and 2th lanes, the difference is small. This may be due to the low expression or low molecular weight of McbA products.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]