Difference between revisions of "Part:BBa K3868103"

 
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===Usage and Biology===
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        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)
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        <img src="https://2021.igem.org/wiki/images/c/c0/T--NNU-China--part-1.png" width="30%" style="float:center">
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:'''Fig 1. The Schematic diagram of pET-AMP-eGFP '''
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K3868103 parameters</partinfo>
 
<partinfo>BBa_K3868103 parameters</partinfo>
 
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Revision as of 11:35, 19 October 2021


pET-Oxysterlin 1-eGFP

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

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