Generation of Nef and GABARAP fusion proteins

The plasmid pDsRed-N2/Nef coding for Nef N-terminally fused to DsRed has been described elsewhere^{34 (link)}. Plasmid peCFP-N1/Nef was generated by subcloning the PCR-amplified Nef coding region from pDsRed-N2/Nef into restriction sites BamHI and HindIII of peCFP-N1 (Clontech). Genes for GABARAP, GABARAPL1, GABARAPL2 and LC3B were subcloned from described GST-fusion plasmids by PCR amplification into the XhoI and BamHI sites of peYFP-C1 (Clontech), yielding peYFP-C1/GABARAP, peYFP-C1/GABARAPL1 and peYFP-C1/GABARAPL2. Sequences of all constructs were confirmed by sequencing.

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Boeske A., Schwarten M., Ma P., Tusche M., Mötter J., Möller C., Neudecker P., Hoffmann S, & Willbold D. (2017). Direct binding to GABARAP family members is essential for HIV-1 Nef plasma membrane localization. Scientific Reports, 7, 5979.

Publication 2017

peCFP-N1 peYFP-C1

Genes Plasmids

Corresponding Organization :

Other organizations : Forschungszentrum Jülich, Heinrich Heine University Düsseldorf

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Variable analysis

independent variables

Plasmid pDsRed-N2/Nef coding for Nef N-terminally fused to DsRed
Plasmid peCFP-N1/Nef generated by subcloning the PCR-amplified Nef coding region from pDsRed-N2/Nef
Genes for GABARAP, GABARAPL1, GABARAPL2 and LC3B subcloned from described GST-fusion plasmids by PCR amplification into the XhoI and BamHI sites of peYFP-C1

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

No positive or negative controls were explicitly mentioned

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