Difference between revisions of "Part:BBa K3712018"

 
 
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<partinfo>BBa_K3712018 short</partinfo>
 
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===USAGE AND BIOLOGY===
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<p>We linked elastin like peptide and BLP-7 by a flexible peptide in order to control the molecular size and gather at the lesion. The function of ELP and BLP-7 has been mentioned before.</p>
  
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We linked elastin like peptide and BLP-7(Optimized) by a flexible peptide in order to control the molecular size and gather at the lesion. The function of ELP and BLP-7 has been mentioned before.
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===RESULTS===
  
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===Usage and Biology===
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<center><img src="https://2021.igem.org/wiki/images/2/2e/T--HUST2-China--the_SDS-PAGE_of_purification.jpg" style="width:646px;height:544px"></center>
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<p><b>Figure 1.</b>SDS-PAGE of Wild-type BLP-7-27 ELP and Optimized BLP-7-27 ELP of purification. lane 1/2 is two-tube stream, lane 3 For Wild-type BLP-7-27 ELP, a thicker band appears near the 15kDa marker; lane 4/5/6 is a three-pipe stream that collects Optimized BLP-7-27 ELP, and a thicker band appears near the 15kDa marker.</p>
  
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<p>After culturing, induction and sterilization, the results of nickel column affinity chromatography show that both of Wild-type BLP-7-27 ELP and Optimized BLP-7-27 ELP elution have single peaks at 80 mM with good symmetry, which preliminarily indicates that the target protein is purified. Then we take the BCA method to determine the concentration (Wild-type BLP-7-27 ELP 1.5mg/ml and Optimized BLP-7-27 ELP 1.2mg/ml) and SDS-PAGE to verify the existence of product.</p>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 13:51, 21 October 2021

Optimized BLP-7-27 ELP

USAGE AND BIOLOGY

We linked elastin like peptide and BLP-7 by a flexible peptide in order to control the molecular size and gather at the lesion. The function of ELP and BLP-7 has been mentioned before.

RESULTS

Part:BBa K3712018

Figure 1.SDS-PAGE of Wild-type BLP-7-27 ELP and Optimized BLP-7-27 ELP of purification. lane 1/2 is two-tube stream, lane 3 For Wild-type BLP-7-27 ELP, a thicker band appears near the 15kDa marker; lane 4/5/6 is a three-pipe stream that collects Optimized BLP-7-27 ELP, and a thicker band appears near the 15kDa marker.

After culturing, induction and sterilization, the results of nickel column affinity chromatography show that both of Wild-type BLP-7-27 ELP and Optimized BLP-7-27 ELP elution have single peaks at 80 mM with good symmetry, which preliminarily indicates that the target protein is purified. Then we take the BCA method to determine the concentration (Wild-type BLP-7-27 ELP 1.5mg/ml and Optimized BLP-7-27 ELP 1.2mg/ml) and SDS-PAGE to verify the existence of product.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 26
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 26
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 26
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 26
  • 1000
    COMPATIBLE WITH RFC[1000]