Difference between revisions of "Part:BBa K3868105"
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pET-24a-SMAP-29 plasmid contains pT7, lacO, SMAP-29, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of SMAP-29 was fused with eGFP in order to characterize the yield of SMAP-29. Moreover, in order to purify the SMAP-29, the enzyme loci of thrombin was inserted between SMAP-29 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-SMAP-29-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of SMAP-29. | pET-24a-SMAP-29 plasmid contains pT7, lacO, SMAP-29, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of SMAP-29 was fused with eGFP in order to characterize the yield of SMAP-29. Moreover, in order to purify the SMAP-29, the enzyme loci of thrombin was inserted between SMAP-29 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-SMAP-29-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of SMAP-29. | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K3868105 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3868105 SequenceAndFeatures</partinfo> | ||
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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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+ | <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> | ||
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===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K3868105 parameters</partinfo> | <partinfo>BBa_K3868105 parameters</partinfo> | ||
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Latest revision as of 11:39, 19 October 2021
pET-SMAP-29-eGFP
pET-24a-SMAP-29 plasmid contains pT7, lacO, SMAP-29, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of SMAP-29 was fused with eGFP in order to characterize the yield of SMAP-29. Moreover, in order to purify the SMAP-29, the enzyme loci of thrombin was inserted between SMAP-29 and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-SMAP-29-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of SMAP-29.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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