Difference between revisions of "Part:BBa K2922030"
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After new molecular cloning experiments, we do Enzyme-Cut identification to certify the plasmid is correct. We use the <i>Eco</i>RI and <i>Pst</i>I to cut the plasmid, then we get the target separate fragment-488bp. (Fig.1) | After new molecular cloning experiments, we do Enzyme-Cut identification to certify the plasmid is correct. We use the <i>Eco</i>RI and <i>Pst</i>I to cut the plasmid, then we get the target separate fragment-488bp. (Fig.1) | ||
<table><tr><th>[[File:TEimm TEkil.png|thumb|300px|Fig.1 The result of this plasmid cut with enzyme <i>Eco</i>RI and <i>Pst</i>I.]]</th><th></table> | <table><tr><th>[[File:TEimm TEkil.png|thumb|300px|Fig.1 The result of this plasmid cut with enzyme <i>Eco</i>RI and <i>Pst</i>I.]]</th><th></table> | ||
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The plasmids are transformed into BL21(DE3).After growing the single colonies,SDS-PAGE protein gel electrophoresis is used to certify the expression of Ekil,then we get the target molecular weight-13kDa.(Fig.2) | The plasmids are transformed into BL21(DE3).After growing the single colonies,SDS-PAGE protein gel electrophoresis is used to certify the expression of Ekil,then we get the target molecular weight-13kDa.(Fig.2) | ||
<table><tr><th>[[File:Eimm protein.png|thumb|300px|Fig.1 The result of the protein molecular weight.]]</th><th></table> | <table><tr><th>[[File:Eimm protein.png|thumb|300px|Fig.1 The result of the protein molecular weight.]]</th><th></table> | ||
Revision as of 14:37, 19 October 2019
T7 promoter-RBS(B0034)_imm(Eimm)
Summary
The usage of T7 promoter and RBS is to highly express the Colicin-E1 immunity protein.
Identification
When we are building this circuit, we are doing the nucleic acid gel electrophoresis experiment to verify. After the circuit is built, we send the plasmid to sequence, and get the correct sequencing. After new molecular cloning experiments, we do Enzyme-Cut identification to certify the plasmid is correct. We use the EcoRI and PstI to cut the plasmid, then we get the target separate fragment-488bp. (Fig.1)
The plasmids are transformed into BL21(DE3).After growing the single colonies,SDS-PAGE protein gel electrophoresis is used to certify the expression of Ekil,then we get the target molecular weight-13kDa.(Fig.2)
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]