Difference between revisions of "Part:BBa K1694010"

 
 
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<partinfo>BBa_K1694010 short</partinfo>
 
<partinfo>BBa_K1694010 short</partinfo>
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<h1>'''Introduction'''</h1>
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Lpp-OmpA protein is the expression system of outer membrane protein, which consists of the 20 amino acids of signal sequence, the 9 N-terminal amino acids of the lipoprotein (Lpp) and the residual 46-159 amino acids of the OmpA.<br>
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The lipoprotein (Lpp) is the most abundant protein on the outer membrane, which possesses the function of targeting to the outer membrane. <br>
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The OmpA domain constitutes eight-stranded β-barrel to construct an anchor on the outer membrane, providing the stable expression of the protein on the outer membrane.<br>
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By taking advantage of efficiently targeting OmpA to the outer membrane, it allows C-terminal fusion of the scFv sequence to be displayed out of the outer mambrane.<br>
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<h2>'''Experiment'''</h2>
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After PCR the DNA sequences from Lpp-OmpA-N , we got the Lpp-OmpA, and then recombined each Lpp-OmpA gene with PSB1C3 backbone and conducted a PCR experiment to check the size of each of the Lpp-OmpA. The DNA sequence length of the Lpp-OmpA is around 400~500 bp. In this PCR experiment, the Lpp-OmpA products size should be near at 650~750 bp. The Fig. showed the correct size of the Lpp-OmpA, and proved that we successfully ligated the Lpp-OmpA sequence onto an ideal backbone.
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[[File:Lpp-OmpA.png|200px|thumb|left|The DNA sequence length of the Lpp-OmpA is around 400~500 bp. In this PCR experiment, the Lpp-OmpA products size should be near at 650~750 bp.]]
  
This part allow us to express scFv on E.coli.
 
  
 
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Latest revision as of 21:42, 22 September 2015

Lpp-OmpA

Introduction

Lpp-OmpA protein is the expression system of outer membrane protein, which consists of the 20 amino acids of signal sequence, the 9 N-terminal amino acids of the lipoprotein (Lpp) and the residual 46-159 amino acids of the OmpA.
The lipoprotein (Lpp) is the most abundant protein on the outer membrane, which possesses the function of targeting to the outer membrane.
The OmpA domain constitutes eight-stranded β-barrel to construct an anchor on the outer membrane, providing the stable expression of the protein on the outer membrane.
By taking advantage of efficiently targeting OmpA to the outer membrane, it allows C-terminal fusion of the scFv sequence to be displayed out of the outer mambrane.

Experiment

After PCR the DNA sequences from Lpp-OmpA-N , we got the Lpp-OmpA, and then recombined each Lpp-OmpA gene with PSB1C3 backbone and conducted a PCR experiment to check the size of each of the Lpp-OmpA. The DNA sequence length of the Lpp-OmpA is around 400~500 bp. In this PCR experiment, the Lpp-OmpA products size should be near at 650~750 bp. The Fig. showed the correct size of the Lpp-OmpA, and proved that we successfully ligated the Lpp-OmpA sequence onto an ideal backbone.

The DNA sequence length of the Lpp-OmpA is around 400~500 bp. In this PCR experiment, the Lpp-OmpA products size should be near at 650~750 bp.


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]