Isolation of cDNA clones for fourteen nuclear-encoded thylakoid membrane proteins

Jochen Tittgen, Jürgen Hermans, Johannes Steppuhn, Thomas Jansen, Christer Jansson, Bertil Andersson, Rachel Nechushtai, Nathan Nelson, Reinhold G. Herrmann*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

72 Scopus citations

Abstract

Spinach cDNA libraries, made from polyadenylated seedling RNA, have been constructed in pBR322 and the expression vector λgt11. Recombinant plasmids or phage for 14 intrinsic and peripheral thylakoid membrane proteins and one stromal protein have been identified. They encode components containing antigenic determinants against the lysine-rich 34 kd, the 23 kd and 16 kd proteins all associated with the water-splitting apparatus of the photosystem II reaction center, the ATP synthase subunits gamma, delta and CFo-II, the Rieske Fe/S protein of the cytochrome b/f complex, subunits 2, 3, 5 and 6 of the photosystem I reaction center, plastocyanin, ferredoxin oxidoreductase, chlorophyll a/b-binding apoproteins of the lightharvesting complex associated with photosystem II, and the small subunit of the stromal enzyme ribulose bisphosphate corboxylase/oxygenase. The cDNA inserts lack complementarity to plastid DNA but hybridize to restricted nuclear DNA as well as to discrete poly A+-mRNA species. The precursor products obtained after translation of hybrid selected RNA fractions in a wheat germ assay are imported and processed by isolated unbroken spinach chloroplasts. The imported components comigrate with the respective authentic proteins.

Original languageAmerican English
Pages (from-to)258-265
Number of pages8
JournalZeitschrift fur Vererbungslehre
Volume204
Issue number2
DOIs
StatePublished - Aug 1986
Externally publishedYes

Keywords

  • Import in organello
  • Spinach
  • Thylakoid membrane polypeptides
  • cDNA expression library
  • transcripts

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