Difference between revisions of "Part:BBa K3868102"

Revision as of 11:34, 19 October 2021

pET-Alloferon-1-eGFP

pET--Alloferon-1-eGFP plasmid contains pT7, lacO,Alloferon-1, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of Alloferon-1 was fused with eGFP in order to characterize the yield of Alloferon-1. Moreover, in order to purify the Alloferon-1, the enzyme loci of thrombin was inserted between Alloferon-1 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-Alloferon-1-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of Alloferon-1.

Sequence and Features

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)