Identification and cloning of the chloroplast gene coding for the large subunit of ribulose-1,5-bisphosphate carboxylase from Chlamydomonas reinhardi.
- 1 August 1977
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 74 (8) , 3193-3197
- https://doi.org/10.1073/pnas.74.8.3193
Abstract
MRNA coding for the large subunit (LS) of ribulose-1,5-bisphosphate carboxylase [3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39] from C. reinhardi was isolated from small polyribosomes immunoadsorbed to column-bound anti-LS antibody. 32P-Labeled LS mRNA was used as a hybridization probe to detect LS genes. The probe hybridized to C. reinhardi chloroplast DNA and at hybridization saturation revealed that there are approximately 75 LS genes per chloroplast. When chloroplast DNA was digested with the restriction endonuclease EcoRI and the fragments were transferred to a nitrocellulose filter, the LS mRNA probe hybridized to a DNA fragment of MW 3.2 .times. 106. This same fragment codes (in part) for 16S and 23S chloroplast rRNA, which are also coded (in part) by fragments of MW 9.0, 2.3 and 0.4 .times. 106. The restriction fragment containing the LS gene was cloned in the Escherichia coli plasmid pMB9.Keywords
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