Team:NU Kazakhstan/Modeling

From 2014.igem.org

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<h3>The model for Nanobodies</h3>
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<h3><center>Nanobodies</center></h3>
<h4>Plasmid design</h4>
<h4>Plasmid design</h4>
<img src="https://static.igem.org/mediawiki/2014/thumb/e/e7/Nb_construct.png/800px-Nb_construct.png" id="img1" width="600" height="320"/>
<img src="https://static.igem.org/mediawiki/2014/thumb/e/e7/Nb_construct.png/800px-Nb_construct.png" id="img1" width="600" height="320"/>
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<p>The construct was synthesized by GenScript company in pUC57 vector</p>
<p>The construct was synthesized by GenScript company in pUC57 vector</p>
<p>We inserted the construct into the pSB1C3 plasmid into the standard restriction sites of EcoRI and PstI</p>
<p>We inserted the construct into the pSB1C3 plasmid into the standard restriction sites of EcoRI and PstI</p>
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<h3>Introducing permanent competence into <i>E. coli</i></h3>
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<h3><center>Introducing permanent competence into <i>E. coli</i></center></h3>
<h4>Making construct</h4>
<h4>Making construct</h4>
<img src="https://static.igem.org/mediawiki/2014/thumb/b/b5/GP16_construct.png/800px-GP16_construct.png" width="600" height="320"/>
<img src="https://static.igem.org/mediawiki/2014/thumb/b/b5/GP16_construct.png/800px-GP16_construct.png" width="600" height="320"/>

Revision as of 16:53, 8 October 2014






Nanobodies

Plasmid design

  • RFP – engineered mutant of red fluorescent protein from Discosoma striata (coral)
  • HlyA- C-terminal signal sequence of alpha-hemolysin

    • The construct was synthesized by GenScript company in pUC57 vector

    We inserted the construct into the pSB1C3 plasmid into the standard restriction sites of EcoRI and PstI

    Introducing permanent competence into E. coli

    Making construct

    The gene for Gp16 ATP-ase protein was ordered from GenScript company.Then, it was combined with the constitutive Anderson promoter + INP, and the constructed part was cloned into standard pSB1C3 plasmid with Circular polymerase extension cloning (CPEC).

    References

    Fraile, S., Muñoz, A., De Lorenzo, V., & Fernández, L. A. (2004). Secretion of proteins with dimerization capacity by the haemolysin type I transport system of Escherichia coli. Molecular microbiology, 53(4), 1109-1121

    Schwartz C, De Donatis GM, Fang H, Guo P. (2013). The ATPase of the phi29 DNA packaging motor is a member of the hexameric AAA+ superfamily. Virology. 443: 20–27.

    Quan J, Tian J. (2009) Circular Polymerase Extension Cloning of Complex Gene Libraries and Pathways. PLoS ONE 4(7): e6441. doi:10.1371/journal.pone.0006441

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