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KMID : 0578319910010040451
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Molecules and Cells
1991 Volume.1 No. 4 p.451 ~ p.457
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Chloroplast Targeting of bacterial β-Glucuronidase with a Pea Transit Peptide in Transgenic Tobacco Plants
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Woo, Jin-Kyeung
Hong, Choo Bong/Lee, June-Seung
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Abstract
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Transit peptides located at the amino termini of the chloroplast precursor proteins are necessary and sufficient components to accomplish import of chloroplast proteins into chloroplasts. We introduced a chimeric gene consisting of a coding sequence of Pisum sativum small subunit of ribulose 1,5-bisphosphate carboxylase/oxygenase (rbcS) transit pep-tide and the gene coding for bacterial 0-glucuronidase (GUS) into tobacco (Nicotiana tabacum L.) cells by Agrobacterium tumefaciens/binary vector system. Transformed tobacco cells with the chimeric gene were in vitro regenerated and the transgenic plants developed were assayed for the expression of GUS and targeting of GUS into chloroplasts by northern blot analysis and measurement of GUS activity. Northern blot analysis confirmed that the fused gene of Gus and the coding sequence for the transit peptide was properly integra- - ted into tobacco chromosome and transcribed. Fractionated chloroplasts from the transgenic tobacco plants showed significant levels of GUS activities. These results indicate that the pea transit peptide is sufficient to transport GUS across the chloroplast envelope into chloroplast in transgenic tobacco plants.
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