Composite

Part:BBa_K3868109

Designed by: Yan Xu   Group: iGEM21_NNU-China   (2021-10-16)


pET-ID13-eGFP

pET-24a-ID13 plasmid contains pT7, lacO, ID13, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of ID13 was fused with eGFP in order to characterize the yield of ID13. Moreover, in order to purify the ID13, the enzyme loci of thrombin was inserted between ID13 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-ID13-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of ID13.


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
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