Recombinant Desmoglein Ectodomains Production

We have previously constructed and expressed the entire extracellular domains of human Dsg1 and Dsg3 using the baculovirus system (Ding et al., 1997 (link); Ding et al., 1999 (link); Flores et al., 2012 (link)). The murine Dsg3 DNA plasmid pEVmod-murineDsg3-His (kind gift of Dr. Masayuki Amagai, Keio University, Tokyo, Japan) was used to generate the mDsg3 recombinant baculovirus using the BacPAK Baculovirus Expression System (Clontech, Mountain View, CA). Soluble ectodomains of hDsg1, hDsg3, and mDsg3 were produced in the baculovirus system and purified by nickel chromatography as described (Flores et al., 2012 (link)).

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Culton D.A., McCray S.K., Park M., Roberts JC J.r., Li N., Zedek D.C., Anhalt G.J., Cowley D., Liu Z, & Diaz L.A. (2015). Mucosal pemphigus vulgaris anti-Dsg3 IgG are pathogenic to the oral mucosa of humanized Dsg3 mice. The Journal of investigative dermatology, 135(6), 1590-1597.

Publication 2015

BacPAK Baculovirus Expression System

Baculovirus Chromatography Human Murine Nickel Plasmid

Corresponding Organization :

Other organizations : University of North Carolina at Chapel Hill, Johns Hopkins University

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Variable analysis

independent variables

Baculovirus system used to generate recombinant mDsg3

dependent variables

Soluble ectodomains of hDsg1, hDsg3, and mDsg3 produced and purified

control variables

Nickel chromatography used to purify the soluble ectodomains

controls

No positive or negative controls were explicitly mentioned.

Annotations

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