Cloned DNA Sequences Complementary to mRNAs Encoding Precursors to the Small Subunit of Ribulose-1,5-bisphosphate Carboxylase and a Chlorophyll a/b Binding Polypeptide

Double-stranded cDNA was synthesized from pea poly(A)-containing mRNA and inserted into the Pst I site of the bacterial plasmid pBR322 by the addition of synthetic oligonucleotide linkers. Bacterial colonies containing recombinant plasmids were detected by hybridization to partially purified mRNAs a...

Full description

Saved in:
Bibliographic Details
Published inProceedings of the National Academy of Sciences - PNAS Vol. 78; no. 12; pp. 7304 - 7308
Main Authors Broglie, Richard, Bellemare, Guy, Bartlett, Sue G., Chua, Nam-Hai, Cashmore, Anthony R.
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences of the United States of America 01.12.1981
National Acad Sciences
Subjects
Biological Sciences: Biochemistry
Chlorophylls
Chloroplasts
Complementary DNA
DNA
Electrophoresis
Gels
Messenger RNA
Peas
Plasmids
Thylakoids
Online AccessGet full text

Cover

More Information
Summary:Double-stranded cDNA was synthesized from pea poly(A)-containing mRNA and inserted into the Pst I site of the bacterial plasmid pBR322 by the addition of synthetic oligonucleotide linkers. Bacterial colonies containing recombinant plasmids were detected by hybridization to partially purified mRNAs and further characterized by cell-free translation of hybridization-selected mRNAs. To confirm the identity of cDNA clones encoding chloroplast polypeptides, we incubated translation products derived from complementary mRNAs with intact chloroplasts in vitro. After uptake, precursor polypeptides were converted to their mature size and identified by fractionation of the chloroplast stroma and thylakoid membranes. By using these procedures, we have isolated and characterized cDNA clones encoding the two major cytoplasmically synthesized chloroplast proteins: the small subunit of ribulose-1,5-bisphosphate carboxylase and a constituent polypeptide (polypeptide 15) of the light-harvesting chlorophyll a/b-protein complex. Similarly, a third cDNA clone was isolated and shown to encode a 22,000-dalton thylakoid membrane polypeptide.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
Present address: Department of Biochemistry, Laval University, Que-bec, PQ, Canada.
Present address: Department of Biochemistry, Louisiana State University, Baton Rouge, LA 70803.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.78.12.7304