Isolation of a cDNA Clone for the γ Subunit of the Chlamydomonas Reinhardtii CF1

  • Lloyd M. Yu
  • Steven M. Theg
  • Bruce R. Selman

Abstract

A cDNA library from Chlamydomonas reinhardtii was probed with antiserum directed against the nuclear encoded γ subunit of the chloroplast H+-translocating ATP synthase (CF0-CF1) of C. reinhardtii. A cDNA was isolated and transcribed in vitro. The transcript was translated in vitro and immunoprecipitated with anti- γ serum to yield a product that co-electrophoresed with the immunoprecipitated product from in vitro translated polyadenylated RNA. These proteins were larger than the mature γ subunit. Thus the γ subunit is synthesized as a precursor of greater molecular weight in C. reinhardtii, as has been suggested for the y subunit from spinach (Nelson et al., 1980).

The precursor protein encoded by this cDNA was imported into pea chloroplasts and processed to a lower molecular weight polypeptide that co-electrophoresed with mature C. reinhardtii γ subunit. The largest cDNA isolated is about the same length as the corresponding mRNA (about 2000 bases). Southern analyses revealed restriction fragment length polymorphisms and that the γ subunit is probably encoded by an intron-containing single copy gene.

Keywords

Formaldehyde Chlorophyll Agar EDTA Agarose 

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Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Lloyd M. Yu
    • 1
  • Steven M. Theg
    • 1
  • Bruce R. Selman
    • 1
  1. 1.Department of Biochemistry, College of Agricultural and Life SciencesUniversity of Wisconsin-MadisonMadisonUSA

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