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Translation of heterologous mRNAs by chloroplast stroma components
Author(s) -
Maiek Ladislav,
Kozieradzki Ivona
Publication year - 1990
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1990.tb09054.x
Subject(s) - chloroplast , polysome , translation (biology) , thylakoid , biochemistry , biology , heterologous , pisum , stroma , protein biosynthesis , amino acid , messenger rna , rna , ribosome , chloroplast stroma , microbiology and biotechnology , gene , immunohistochemistry , immunology
In vitro translation of exogenous mRNAs has been difficult to achieve using chloroplast components. An in vitro protein synthesis system is described, based on a pea ( Pisum sativum L, cv. Spring) chloroplast stroma 30000 g supernatant, which was capable of translating polyuridylie acid and MS2 phage RNA into corresponding proteins. The kinetics of label incorporation and fluorograms of unique translation products are presented. The unusual steps which may have contributed to successful translation include the following: the removal of exogenous unlabeled amino acids from the incubation mixture, ultrafiltration of the stromal supernatant and the removal of thylakoid‐bound polyribosomes. The system could be further developed to study translational control of gene expression in the chloroplasts.