Cloning of cDNA encoding the precursor to the small subunit of ribulose-1,5-bisphosphate carboxylase in pea (Pisum sativum)
1989
Chang, M.W. | Koo, Y.B. (Yeongnam Univ., Kyonsan (Korea R.). Dept. of Biology) | Kim, H.J. (Ajou Univ., Suwon (Korea R.). Dept. of Biology)
Polysomal polyadenylated mRNAs which were purified from pea leaves were fractionated by sucrose grandient sedimentation. Fractions corresponding to the peak at 11.5S were found to contain mostly mRNA encoding the precursor polypeptide to the small subunit of ribulose bisphosphate carboxylase (rbcS) by in vitro translation in wheat germ extract. Double-stranded cDNA which was synthesized from the 11.5S mRNA was cloned into Hind III site of plasmid pBR 325. A cDNA clone, H24, was identified to code for rbcS. In vitro translation product of the hybridization-selected mRNA was molecular weight 20,000, presumably the precursor of rbcS. The nucleotide sequences of the H24 showed almost complete homology with the sequences encoding the transit peptide of the rbcS-3A gene which was reported by Fluhr et al. (1986)
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