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Soluble full-length expression and characterization of snRNP protein U1-68/70K.
Richter, Carolin; Simon, Thomas; Asen, Iris; Brenner-Weiss, Gerald; Hubbuch, Jürgen.
Afiliación
  • Richter C; DIARECT AG, Freiburg, Germany; Karlsruhe Institute of Technology, Institute of Process Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Engler-Bunte-Ring 1, 76131 Karlsruhe, Germany. Electronic address: Carolin.Richter@kit.edu.
  • Simon T; DIARECT AG, Freiburg, Germany. Electronic address: Thomas.Simon@diarect.com.
  • Asen I; DIARECT AG, Freiburg, Germany. Electronic address: Iris.Asen@diarect.com.
  • Brenner-Weiss G; Karlsruhe Institute of Technology, Campus Nord, Institute of Functional Interfaces, Analytical Biochemistry, Eggenstein-Leopoldshafen, Germany. Electronic address: Gerald.Brenner-Weiss@kit.edu.
  • Hubbuch J; Karlsruhe Institute of Technology, Institute of Process Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Engler-Bunte-Ring 1, 76131 Karlsruhe, Germany. Electronic address: Juergen.Hubbuch@kit.edu.
Protein Expr Purif ; 104: 65-70, 2014 12.
Article en En | MEDLINE | ID: mdl-25201698
The autoantigen U1-68/70K is the dominant diagnostic marker in Mixed Connective Tissue Disease (MCTD) that until recently could not be expressed in its full-length form (Northemann et al., 1995, [16]). Using cell-free expression screening, we successfully produced the snRNP protein U1-68/70K in a soluble full-length form in Escherichia coli cells. The protein length and identity was determined by Western Blot and MS/MS analysis. Additionally, its reactivity in the autoimmune diagnostic was confirmed. Establishment of a cell-free expression system for this protein was important for further elucidation of protein expression properties such as the cDNA construct, expression temperature and folding properties; these parameters can now be determined in a fast and resource-conserving manner.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Autoantígenos / Ribonucleoproteína Nuclear Pequeña U1 Límite: Humans Idioma: En Revista: Protein Expr Purif Asunto de la revista: BIOLOGIA MOLECULAR Año: 2014 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Autoantígenos / Ribonucleoproteína Nuclear Pequeña U1 Límite: Humans Idioma: En Revista: Protein Expr Purif Asunto de la revista: BIOLOGIA MOLECULAR Año: 2014 Tipo del documento: Article Pais de publicación: Estados Unidos