1. A Single Common Protocol for the Expression and Purification of Soluble Mammalian DSPs from Escherichia coli.
- Author
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Stepanyants N, Macdonald PJ, Madan Mohan P, and Ramachandran R
- Subjects
- Animals, Escherichia coli metabolism, Plasmids genetics, Solubility, Transformation, Bacterial, Desmoplakins genetics, Desmoplakins isolation & purification, Escherichia coli genetics, Gene Expression, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins isolation & purification
- Abstract
Mammalian DSPs have been historically isolated either from native tissue sources or from transfected insect cell cultures via time-consuming and cumbersome protocols often yielding protein of variable quality and quantity. A facile and highly reproducible alternative methodology involving the heterologous expression and purification of soluble mammalian DSPs from E. coli, which yields highly active and functional protein of a uniform quality and quantity, free of spurious posttranslational modifications inherent to mammalian and insect cell expression systems, is described in this chapter.
- Published
- 2020
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