Difference between revisions of "Team:Freiburg/Project/pRIG15 17"
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| − | The <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Project/Diseases#AIDS" title="aids">Human | + | The <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Project/Diseases#AIDS" title="aids">Human Immunodeficiency Virus (HIV)</a> causes acquired immunodeficiency syndrome, AIDS, which leads to major impairments of the immunsystem. <sup><a class="fn_top" href="#fn__1" id="fnt__1" name="fnt__1">1)</a></sup> For our experiments we used a sequence coding for a recombinant protein that contains six epitopes from different proteins of the HI Virus (HIV-1-trans-activating (tat) encoding region, one epitope of the reverse transcriptase, one of the p24 protein, one of the envelope protein gp41, one of gp120).<sup><a class="fn_top" href="#fn__2" id="fnt__2" name="fnt__2">2)</a></sup> |
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Revision as of 16:23, 16 September 2015
pRIG15_17
The Human Immunodeficiency Virus (HIV) causes acquired immunodeficiency syndrome, AIDS, which leads to major impairments of the immunsystem. 1) For our experiments we used a sequence coding for a recombinant protein that contains six epitopes from different proteins of the HI Virus (HIV-1-trans-activating (tat) encoding region, one epitope of the reverse transcriptase, one of the p24 protein, one of the envelope protein gp41, one of gp120).2)
To insert the sequence for HIV tat/pol/gag/env into pSB1C3 we designed Gibson primers with compatible overhangs that also included the start codon ATG. This fragment was amplified via PCR (Link zum Labjournal-Eintrag) and then assembled with the digested pSB1C3 backbone using Gibson assembly. To prove correct insertion of our fragment we did a test digest and sent the whole plasmid for sequencing.
We inserted the sequence coding for HIV tat/pol/gag/env into pET_22b+ for overexpression in E.coli. We could show interaction of the HIV tat/pol/gag/env antigen with a polyclonal anti-HIV-1 P24 antibody (Fig. 2)
Get the sequence in GeneBank format: BBa_K1621004.gb.
