Difference between revisions of "Part:BBa K3722010"
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | < | + | <Biology |
| + | SttH can substitute the 6-Br-Trp into a Bromine atom. | ||
| + | Fre is a NAD(P)H-flavin reductase from E. coli . It is highly expressed in a soluble form in E. coli and can regenerate FADH2 efficiently. | ||
| + | |||
| + | Usage | ||
| + | Here, we use BBa_K3722006 to increase the solubility of SttH as a fusion enzyme. We transformed the composite part BBa_K3722010 into E. coli BL21(DE3) to improve the yield of 6-chloro-Trp. | ||
| + | |||
| + | Characterization | ||
| + | The proof of expression | ||
| + | Based on literature review and pre-experiment, we found it was easy to express in the form of inclusion body. So we induced BBa_K3722006 at different temperatures to find the optimum conditions. We determined the expression level through SDS-PAGE. the experimental results were shown below.附上蛋白表达图> | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K3722010 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3722010 SequenceAndFeatures</partinfo> | ||
Revision as of 12:08, 21 October 2021
T7_Promoter-RBS-Fre-Linker-SttH-T7_Terminator
Fre is a unique N-terminal tag that efficiently enhanced SttH activity by increasing solubility as well as cofactor supply, the improvement might be caused by an enhanced electron transfer efficiency due to the prox-imity effect of the fusion of Fre and SttH, and greatly improved the yield and productivity of bromination.