BMB Reports

Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Isolation and Characterization of Exogenously Expressed Calmodulin from Endogenous Tobacco Calmodulin by Anion-exchange Fast Protein Liquid Chromatography

  • Oh, Suk-Heung (Department of Biotechnology, Woosuk University) ;
  • Cha, Youn-Soo (Department of Biochemistry, Chonbuk National University Medical School) ;
  • Lee, Tae-Kyoo (Department of Food Science & Technology, Woosuk University)
  • Published : 1995.07.31
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Abstract

A Mono Q HR 5/5 anion-exchange column with a FPLC system was used to separate exogenously expressed calmodulin from endogenous tobacco calmodulins. Transgenic tobacco calmodulins were purified by fractionation with ammonium sulfate, precipitation with sulfuric acid and hydrophobic chromatography on phenyl-Sepharose CL-4B. The purified calmodulins were chromatographed in the FPLC using the column. This method was selected because of the slight differences in the net charge of foreign and endogenous plant calmodulins due to amino acid sequence differences. By this approach, the exogenously expressed calmodulin was isolated from endogenous tobacco calmodulins. The isolated calmodulin was characterized by amino acid composition analysis as well as methylation analysis.

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