Difference between revisions of "Part:BBa K3868108"
 
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Latest revision as of 11:49, 19 October 2021


pET-PEW300-eGFP

pET-24a-PEW300 plasmid contains pT7, lacO, PEW300, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of PEW300 was fused with eGFP in order to characterize the yield of PEW300. Moreover, in order to purify the PEW300, the enzyme loci of thrombin was inserted between PEW300 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-PEW300-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of PEW300.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE 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