Recombinant Tau Isoform Expression

Human tau isoforms were cloned into the pRK172 bacterial expression vector using NdeI and EcoRI restriction sites as previously described [27 (link), 31 (link), 75 (link)]. Recombinant tau isoforms were expressed in Escherichia coli BL21 (New England Biolabs, Ipswich, MA) and purified. In brief, expression was induced by isopropyl-β-D-1-thiogalactopyranoside (IPTG) and high-salt 100 °C heat stable bacterial protein lysates were purified by MonoS cation exchange chromatography.

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Paterno G., Bell B.M., Gorion K.M., Prokop S, & Giasson B.I. (2022). Reassessment of Neuronal Tau Distribution in Adult Human Brain and Implications for Tau Pathobiology. Acta Neuropathologica Communications, 10, 94.

Publication 2022

Escherichia coli BL21

Bacterial Bacterial lysates Chromatography Ecori Escherichia coli Human Iptg Isoforms Monos Protein Salt Vector

Corresponding Organization :

Other organizations : University of Florida

Top 5 similar protocols

Variable analysis

independent variables

Tau isoforms

dependent variables

Recombinant tau isoform expression
Tau protein purification

control variables

Escherichia coli BL21 bacterial expression system
IPTG for induction of protein expression
High-salt 100 °C heat treatment for bacterial protein lysate preparation
MonoS cation exchange chromatography for protein purification

controls

Positive control: Induction of recombinant tau isoform expression in E. coli BL21 using IPTG
Negative control: Not explicitly mentioned

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