Cloning and Expression of Par Complex Components

GST-, MBP-, and his-tagged Par-3 constructs, GST-aPKC PBM, and his aPKC kinase domain-PBM (residues 259–606) were cloned as previously described (10 (link)) using Gibson cloning (New England Biolabs), Q5 mutagenesis (New England Biolabs), or traditional methods. In addition to an N-terminal MBP tag, the Par-3 PDZ1-APM (residues 309–987) construct also contained a C-terminal his-tag. Par complex components (aPKC and his-Par-6) were cloned into pCMV (human cytomegalovirus) as previously described (10 (link), 23 (link)). Please see the Key resources table for additional information on specific constructs.

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Penkert R.R., Vargas E, & Prehoda K.E. (2022). Energetic determinants of animal cell polarity regulator Par-3 interaction with the Par complex. The Journal of Biological Chemistry, 298(8), 102223.

Publication 2022

Gibson cloning Q5 mutagenesis

Human cytomegalovirus Kinase Mutagenesis

Corresponding Organization : University of Oregon

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Variable analysis

independent variables

GST-, MBP-, and his-tagged Par-3 constructs
GST-aPKC PBM
His aPKC kinase domain-PBM (residues 259–606)

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive controls: Not mentioned
Negative controls: Not mentioned

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