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Nucleic Acids Research, 1992, Vol. 20, No. 4 689-697
© 1992

Articles

Nuclear-encoded chloroplast ribosomal protein L12 of Nicotians tabacum: characterization of mature protein and isolation and sequence analysis of cDNA clones encoding its cytoplasmic precursor

Gasmalla A. EIhag1, Frank J. Thomas1,+, Thomas P. McCreery1 and Don P. Bourque1,2,*

1Department of Biochemistry, University of Arizona Tucson, AZ 85721,USA 2Department of Molecular and Cellular Biology, University of Arizona Tucson, AZ 85721, USA

*To whom correspondence should be addressed

Received December 11, 1991. Revised January 22, 1992. Accepted January 22, 1992.

Poly(A)+ mRNA isolated from Nicotians tabacum (cv. Petite Havana) leaves was used to prepare a cDNA library in the expression vector {lambda}gt11. Recombinant phage containing cDNAs coding for chloroplast ribosomal protein L12 were identified and sequenced. Mature tobacco L12 protein has 44% amino acid identity with ribosomal protein L7/L12 of Escherichla coli. The longest L12 cDNA (733 nucleotides) codes for a 13,823 molecular weight polypeptide with a transit peptide of 53 amino acids and a mature protein of 133 amino acids. The transit peptide and mature protein share 43% and 79% amino acid identity, respectively, with corresponding regions of spinach chloroplast ribosomal protein L12. The predicted amino terminus of the mature protein was confirmed by partial sequence analysis of HPLC-purified tobacco chloroplast ribosomal protein L12. A single L12 mRNA of about 0.8 kb was detected by hybridization of L12 cDNA to poly(A)+ and total leaf RNA. Hybridization patterns of restriction fragments of tobacco genomlc DNA probed with the L12 cDNA suggested the existence of more than one gene for ribosomal protein L12. Characterization of a second cDNA with an identical L12 coding sequence but a different 3'-non-coding sequence provided evidence that at least two L12 genes are expressed in tobacco.

+Present address: Laboratoire de Physiologie Cellulaire Vegetale, Centre National de la Recherche Seientifique, Universite Scientifique et Medicate de Grenoble, F-38041 Grenoble-Cedex, France


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[Abstract] [Full Text] [PDF]


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