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Expression of the atpE Gene Coding for the epsilon Subunit of Maize Chloroplast Coupling Factor
Jin Shi1, Jian-Mian Wei, Yun-Gang Shen
1Shanghai Institute of Plant Physiology, Academia Sinica, Shanghai 200032, China.
Abstract:
The entire atpE gene of the maize chloroplast coupling factor was inserted into the polylinker region of vectors pJLA505 and pWA to form recombinant plasmids pJLA505-atpE and pWA-atpE respectively. These expression plasmids were transformed into E. coli NM522 which induced at 42 degrees. By the analysis of SDS-PAGE, the expressed product of interest was observed to account fore more than 3o% of total E. coliproteins. The identification of the expressed product demonstrated that its immunological specificity was well retained. The antiserum cross-reacted with the expressed epsilon protein and CF(1)-epsilon protein of spinach and produced precipitin lines on Ouchterlony immunodiffusion test. The expressed product aggregated insolubly as the inclusion body and was purified to over 80% purity. The purified product had the same function as that of the native epsilon subunit.