Generation and Characterization of Mutant GPR35 Constructs

Generation of FLAG-hGPR35a-eYFP, hGPR35a-HA, FLAG-mGPR35-eYFP, and mGPR35-HA have been described previously (35 (link), 36 (link)). The Stratagene Quik Change method (Stratagene, Agilent Technologies) was used to introduce alterations into each of the above constructs to produce both point mutants and phospho-deficient variants. Primers utilised for mutagenesis were from MWG Operon. Sequencing was carried out to confirm the introduction of the alterations.

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Ganguly A., Quon T., Jenkins L., Joseph B., Al-awar R., Chevigne A., Tobin A.B., Uehling D.E., Hoffmann C., Drube J, & Milligan G. (2023). G protein–receptor kinases 5/6 are the key regulators of G protein–coupled receptor 35–arrestin interactions. The Journal of Biological Chemistry, 299(10), 105218.

Publication 2023

Stratagene Quik Change method

Mutagenesis Operon Primers

Corresponding Organization : University of Glasgow

Other organizations : Ontario Institute for Cancer Research, Luxembourg Institute of Health, Friedrich Schiller University Jena

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Variable analysis

independent variables

Alterations introduced into each of the above constructs to produce both point mutants and phospho-deficient variants

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive controls: Not specified
Negative controls: Not specified

Annotations

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