Difference between revisions of "Part:BBa K3868107"

 
 
Line 5: Line 5:
 
pET-24a-SPINK9 plasmid contains pT7, lacO, SPINK9, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of SPINK9 was fused with eGFP in order to characterize the yield of SPINK9. Moreover, in order to purify the SPINK9, the enzyme loci of thrombin was inserted between SPINK9 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-SPINK9-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of SPINK9.
 
pET-24a-SPINK9 plasmid contains pT7, lacO, SPINK9, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of SPINK9 was fused with eGFP in order to characterize the yield of SPINK9. Moreover, in order to purify the SPINK9, the enzyme loci of thrombin was inserted between SPINK9 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-SPINK9-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of SPINK9.
  
−
<!-- Add more about the biology of this part here
 
−
===Usage and Biology===
 
  
−
<!-- -->
 
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3868107 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3868107 SequenceAndFeatures</partinfo>
  
  
 +
 +
===Usage and Biology===
 +
 +
&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)
 +
 +
<html>
 +
<div align="center">
 +
    <figure>
 +
        <img src="https://2021.igem.org/wiki/images/c/c0/T--NNU-China--part-1.png" width="30%" style="float:center">
 +
        <figcaption>
 +
        <p style="font-size:1rem">
 +
        </p>
 +
        </figcaption>
 +
    </figure>
 +
</div>
 +
</html>
 +
<div align="center">
 +
:'''Fig 1. The Schematic diagram of pET-AMP-eGFP '''
 +
</div>
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K3868107 parameters</partinfo>
 
<partinfo>BBa_K3868107 parameters</partinfo>
 
<!-- -->
 
<!-- -->

Latest revision as of 11:46, 19 October 2021


pET-SPINK9-eGFP

pET-24a-SPINK9 plasmid contains pT7, lacO, SPINK9, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of SPINK9 was fused with eGFP in order to characterize the yield of SPINK9. Moreover, in order to purify the SPINK9, the enzyme loci of thrombin was inserted between SPINK9 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-SPINK9-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of SPINK9.


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