Cloning and Expressing Human α1 and α2

DNAs encoding full-length human α1 and α2 were amplified with primers designed to encode a 5′ Kpn1 site, and a 3′ FLAG-tag followed by an Xho1 site. The resulting PCR products were cloned into the pcDNA5/FRT plasmid (Invitrogen). Stable cell lines were generated and cultured as described previously [19 (link)].

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Hawley S.A., Ross F.A., Gowans G.J., Tibarewal P., Leslie N.R, & Hardie D.G. (2014). Phosphorylation by Akt within the ST loop of AMPK-α1 down-regulates its activation in tumour cells. Biochemical Journal, 459(Pt 2), 275-287.

Publication 2014

pcDNA5/FRT plasmid

Cell lines Dnas Human Plasmid Primers

Corresponding Organization :

Other organizations : University of Dundee

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Variable analysis

independent variables

Insertion of DNA encoding full-length human α1 and α2 into pcDNA5/FRT plasmid

dependent variables

Generation of stable cell lines expressing human α1 and α2 proteins

control variables

Cell culture conditions as described previously [19]

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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