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Immunoblot analysis of the expression of genes for barley rubisco activase inE. coli

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Abstract

Rubisco activity during photosynthesis is regulated by the rubisco activase, which facilitates the dissociation of RuBP and other inhibitory sugar phosphates from the active site of rubisco in an ATP-dependent reaction. In this paper, barleyRca genes (RcaA1,RcaA2 andRcaB) were expressed inE. coli and the activity of rubisco activase expressed was assayed biochemically by chromatography. Then the protein was identified electrophoretically by SDS-PAGE and detected immunologically by Western blot analysis using polyclonal antibodies raised against the kidney bean rubisco activase as probe. The band pattern of purified proteins on the polyacrylamide gel showed two polypeptides of 46 kD and 42 kD. Anti-rubisco activase antibodies reacted specifically with both polypeptides of 46 kD and 42 kD present in the crude extracts ofE. coli transformants. Therefore, it was found that the genes of barley rubisco activase was successfully expressed inE. coli as active forms of 46 kD and 42 kD.

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Authors and Affiliations

  1. Department of Biology, Keimyung University, 704-701, Taegu, Korea

    Kwang Soo Ron, Mo Sun Kwon, Young Hwan Do, Seung Dal Song, Hwa Sook Chung & Jong Suk Song

  2. Department of Biology, Kyungpook National University, 702-701, Taegu, Korea

    Seung Dal Song

  3. Department of Biological Education, Kyungpook National University, 702-701, Taegu, Korea

    Hwa Sook Chung

  4. Department of Biology, Andong National University, 760-749, Andong, Korea

    Jong Suk Song

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  1. Kwang Soo Ron
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  2. Mo Sun Kwon
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  4. Seung Dal Song
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Ron, K.S., Kwon, M.S., Do, Y.H. et al. Immunoblot analysis of the expression of genes for barley rubisco activase inE. coli . J. Plant Biol. 41, 233–239 (1998). https://doi.org/10.1007/BF03030258

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  • DOI: https://doi.org/10.1007/BF03030258

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