Composite

Part:BBa_K3868100

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


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


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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