Bacterial strains and plasmids

Yasha Butt; Ramin Sakhtemani; Rukshana Mohamad-Ramshan; Michael S. Lawrence; Ashok S. Bhagwat

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Bacterial strains and plasmids

YB Yasha Butt

RS Ramin Sakhtemani

RM Rukshana Mohamad-Ramshan

ML Michael S. Lawrence

AB Ashok S. Bhagwat

This method is extracted from research article: Nat Commun, Mar 2024

Distinguishing preferences of human APOBEC3A and APOBEC3B for cytosines in hairpin loops, and reflection of these preferences in APOBEC-signature cancer genome mutations

DOI: 10.1038/s41467-024-46231-w

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E. coli strains including BH214 (GM31 ung^- mug^-) and BL21(DE3) were respectively used for the expression of either A3B-CTD or A3B-full, and the purification of A3B-CTD and A3A proteins. Human A3B-CTD and A3B-full were separately cloned into pASK-IBA5C plasmid vector (IBA LifeSciences) using Gibson cloning kit (Thermo Fisher Scientific). The clones were confirmed using colony PCR followed by the digestion of plasmid DNA using appropriate restriction enzymes and by Sanger sequencing (Genomics Core, Michigan State University). A3B-CTD was also cloned into pET28a plasmid vector for protein purification purposes and its sequence was also confirmed using Sanger sequencing.

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