Difference between revisions of "Part:BBa K2631000"
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===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K2631000 parameters</partinfo> | <partinfo>BBa_K2631000 parameters</partinfo> | ||
| + | </h1>Expression of MBP-ADTZ</h3> | ||
| + | Firstly the vector pMAL-c5x-ADTZ was transformed into E.coli BL21 GOLD DE(3). Then an overnight culture of the bacteria was inoculated into LB broth containing 2g/L glucose and 100mg/L ampicillin. After 2h culture at 37 oC and 200rpm, recombinant protein production was induced by addition of 0.3 mM IPTG at 20oC and 200rpm for 24h. The cell was collected by centrifugation. The pellet was resuspended in H2O about an OD600 of 30. Then the cell was disrupted by sonication. The results showed that most of highly expressed MBP-ADTZ(119kDa) was soluble(Fig. 1). | ||
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Revision as of 05:49, 9 October 2018
ADTZ Enzyme
ADTZ, which is also called aflatoxin oxidase (AFO), is the first enzyme identified to bed able to degrade AFB1. It can be isolated from intracellular extracts and show a strong affinity with AFB1. As a potential degradation enzyme candidate in our project. In order to express soluble ADTZ in E．coli，the DNA coding ADTZ was inserted into the vector pMAL-c5x to produce the fusion protein of maltose-binding protein (MBP) and ADTZ.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1556
Illegal BglII site found at 1979 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]