Difference between revisions of "Part:BBa K1694013"
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<h1>'''Introduction:'''</h1>
 
<h1>'''Introduction:'''</h1>
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[[File:ompascfv.png|200px|thumb|left|'''fig.1''' ompascfv]]
 
 
To display the antibody outside the E.coli, we used Lipoprotein-Outer membrane protein A (Lpp-OmpA). According to the paper reference [1], We chose the first 9 amino acid of Lpp, and the 46~159 amino acid of OmpA.
 
To display the antibody outside the E.coli, we used Lipoprotein-Outer membrane protein A (Lpp-OmpA). According to the paper reference [1], We chose the first 9 amino acid of Lpp, and the 46~159 amino acid of OmpA.
 
<br>
 
<br>
 
In order to change the scFv parts easily, we added a NcoI restriction site between OmpA and scFv so that we can change vario us scFv DNA sequence using the NcoI restriction enzyme.
 
In order to change the scFv parts easily, we added a NcoI restriction site between OmpA and scFv so that we can change vario us scFv DNA sequence using the NcoI restriction enzyme.
 +
 +
[[File:ompascfv.png|200px|thumb|center|'''Fig.1''' ompascfv]]
 +
  
 
<h1>'''Experiment:'''</h1>
 
<h1>'''Experiment:'''</h1>
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After receiving the DNA sequences from the gene synthesis company, we recombined each OmpA-N-scFv gene to PSB1C3 backbones and conducted a PCR experiment to check the size of each of the OmpA-N-scFvs. The DNA sequence length of the OmpA-N-scFvs are around 1000~1200 bp. In this PCR experiment, the OmpA-N-scFv products size should be near at 1200~1400 bp. The Fig. showed the correct size of the scFv, and proved that we successful ligated the scFv sequence onto an ideal backbone.
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[[File:OB.png|200px|thumb|left|The PCR result of the Lpp-OmpA-N+scFv. The DNA sequence length is around 1000~1200 bp, so the PCR products should appear at 1200~1400 bp.]]
 +
 
 +
After receiving the DNA sequences from the gene synthesis company, we recombined each OmpA-N-scFv gene to PSB1C3 backbones and conducted a PCR experiment to check the size of each of the OmpA-N-scFv. The DNA sequence length of the OmpA-N-scFv is around 1000~1200 bp. In this PCR experiment, the OmpA-N-scFv products size should be near at 1200~1400 bp. The Fig.3 showed the correct size of the OmpA-N-scFv, and proved that we successful ligated the scFv sequence onto an ideal backbone.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 14:50, 19 September 2015

OmpA-scFv(Anti-VEGF)

Introduction:

To display the antibody outside the E.coli, we used Lipoprotein-Outer membrane protein A (Lpp-OmpA). According to the paper reference [1], We chose the first 9 amino acid of Lpp, and the 46~159 amino acid of OmpA.
In order to change the scFv parts easily, we added a NcoI restriction site between OmpA and scFv so that we can change vario us scFv DNA sequence using the NcoI restriction enzyme.

Fig.1 ompascfv


Experiment:

The PCR result of the Lpp-OmpA-N+scFv. The DNA sequence length is around 1000~1200 bp, so the PCR products should appear at 1200~1400 bp.

After receiving the DNA sequences from the gene synthesis company, we recombined each OmpA-N-scFv gene to PSB1C3 backbones and conducted a PCR experiment to check the size of each of the OmpA-N-scFv. The DNA sequence length of the OmpA-N-scFv is around 1000~1200 bp. In this PCR experiment, the OmpA-N-scFv products size should be near at 1200~1400 bp. The Fig.3 showed the correct size of the OmpA-N-scFv, and proved that we successful ligated the scFv sequence onto an ideal backbone.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 388
  • 1000
    COMPATIBLE WITH RFC[1000]