Difference between revisions of "Part:BBa K3868100"
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Latest revision as of 11:31, 19 October 2021
pET-MME-eGFP
pET--MME-eGFP plasmid contains pT7, lacO, MME, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of MME was fused with eGFP in order to characterize the yield of MME. Moreover, in order to purify the MME , the enzyme loci of thrombin was inserted between MME and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-MME-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of MME.
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]
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