Part:BBa_K3868101
pET-Fa-AMP1-eGFP
pET-24a-Fa-AMP1 plasmid contains pT7, lacO, Fa-AMP1, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of Fa-AMP1 was fused with eGFP in order to characterize the yield of Fa-AMP1. Moreover, in order to purify the Fa-AMP1, the enzyme loci of thrombin was inserted between Fa-AMP1 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-Fa-AMP1-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of Fa-AMP1.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 111
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 111
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 111
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 111
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
Based on the plasmid of pET-24a. The C-terminal of Antibacterial peptides(AMP) was fused with eGFP in order to characterize the yield of AMP. The pET-24a was provided by our PI. Moreover, in order to purify the AMP, the enzyme loci of thrombin was inserted between AMP and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-AMP-eGFP. As a result, the pET-AMP-eGFP plasmids could not only indicate the expression level of AMPs by the fluorescence intensity of eGFP, but also be good for later purification and separation with His-Tag and the enzyme loci of thrombin. (Fig. 1)
- Fig 1. The Schematic diagram of pET-AMP-eGFP
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