Difference between revisions of "Part:BBa K1694023"
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<h1>'''Experiment'''</h1>
 
<h1>'''Experiment'''</h1>
 
<p style="font-size:120%">'''1.Cloning'''</p>
 
<p style="font-size:120%">'''1.Cloning'''</p>
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[[File:PROVEGF.png|200px|thumb|left|The PCR result of the Pcons+B0034+Lpp-OmpA-N+scFv. The DNA sequence length is around 1100~1300 bp, so the PCR products should appear at 1300~1500 bp.]]  
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[[File:PROBPCR.png|200px|thumb|left|'''Fig.3'''The PCR result of the Pcons+B0034+Lpp-OmpA-N+scFv. The DNA sequence length is around 1100~1300 bp, so the PCR products should appear at 1300~1500 bp.]]  
 
After assemble the DNA sequences from the basic parts, we recombined each Pcons+RBS+Lpp-OmpA-N+scFv gene to PSB1C3 backbones and conducted a PCR experiment to check the size of each of the parts. The DNA sequence length of these parts is around 1100~1300 bp. In this PCR experiment, the scFv products size should be near at 1300~1500 bp. The Fig. 3 showed the correct size of the scFv, and proved that we successful ligated the scFv sequence onto an ideal backbone.
 
After assemble the DNA sequences from the basic parts, we recombined each Pcons+RBS+Lpp-OmpA-N+scFv gene to PSB1C3 backbones and conducted a PCR experiment to check the size of each of the parts. The DNA sequence length of these parts is around 1100~1300 bp. In this PCR experiment, the scFv products size should be near at 1300~1500 bp. The Fig. 3 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:PROBplate.png|600px|thumb|center|'''Fig.4'''Pcons+B0034+Lpp-OmpA-N+scFv(Anti-VEGF)]]
  
  

Revision as of 04:20, 20 September 2015

Pcons+B0034+Lpp-OmpA-N+scFv(Anti-VEGF)

Introduction:

Fig.1 Pcons+B0034+Lpp-OmpA-N+scFv(Anti-VEGF)

By ligating the constitutive promoter (BBa_J23101), strong ribosome binding site (BBa_B0034) and Lpp-OmpA-scFv, we were able to display scFv(Bevacizumab) outside the E.coli cell membrane.

This year we want to provide a customized platform. We provide two libraries of Pcon+RBS+OmpA-scFv and Pcons+RBS+Fluorescence+Ter into E. coli. Therefore, our customers can choose any scfv and any fluorescence protein. Our team will then co-transform the two plasmids, which helps us tailor our product to the wishes of our customers.

Experiment

1.Cloning

Fig.3The PCR result of the Pcons+B0034+Lpp-OmpA-N+scFv. The DNA sequence length is around 1100~1300 bp, so the PCR products should appear at 1300~1500 bp.

After assemble the DNA sequences from the basic parts, we recombined each Pcons+RBS+Lpp-OmpA-N+scFv gene to PSB1C3 backbones and conducted a PCR experiment to check the size of each of the parts. The DNA sequence length of these parts is around 1100~1300 bp. In this PCR experiment, the scFv products size should be near at 1300~1500 bp. The Fig. 3 showed the correct size of the scFv, and proved that we successful ligated the scFv sequence onto an ideal backbone.


Fig.4Pcons+B0034+Lpp-OmpA-N+scFv(Anti-VEGF)


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 451
  • 1000
    COMPATIBLE WITH RFC[1000]