Difference between revisions of "Part:BBa K3868101"

 
 
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pET-24a-Fa-AMP1 plasmid contains pT7, lacO, Fa-AMP1, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of Fa-AMP1 was fused with eGFP in order to characterize the yield of Fa-AMP1. Moreover, in order to purify the Fa-AMP1, the enzyme loci of thrombin was inserted between Fa-AMP1 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-Fa-AMP1-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of Fa-AMP1.
 
pET-24a-Fa-AMP1 plasmid contains pT7, lacO, Fa-AMP1, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of Fa-AMP1 was fused with eGFP in order to characterize the yield of Fa-AMP1. Moreover, in order to purify the Fa-AMP1, the enzyme loci of thrombin was inserted between Fa-AMP1 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-Fa-AMP1-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of Fa-AMP1.
  
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===Usage and Biology===
 
  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3868101 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3868101 SequenceAndFeatures</partinfo>
  
  
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===Usage and Biology===
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;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_K3868101 parameters</partinfo>
 
<partinfo>BBa_K3868101 parameters</partinfo>
 
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Latest revision as of 11:32, 19 October 2021


pET-Fa-AMP1-eGFP

pET-24a-Fa-AMP1 plasmid contains pT7, lacO, Fa-AMP1, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of Fa-AMP1 was fused with eGFP in order to characterize the yield of Fa-AMP1. Moreover, in order to purify the Fa-AMP1, the enzyme loci of thrombin was inserted between Fa-AMP1 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-Fa-AMP1-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of Fa-AMP1.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 111
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 111
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 111
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 111
  • 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