Transient expression of photosynthetic genes in transfected albinoid petunia protoplasts and correct processing of newly synthesized chloroplast-destined polypeptides
作者： Shimsbon Broido , Abraham Loyter , Alexander Vainstein
DOI: 10.1111/J.1399-3054.1993.TB05497.X
关键词: Gene expression 、 Molecular biology 、 RuBisCO 、 Western blot 、 Biology 、 Cauliflower mosaic virus 、 Expression vector 、 Chloroplast 、 Petunia 、 Tobacco mosaic virus
摘要: Protoplasts prepared from cultured albinoid cells of petunia do not express photosynthetic genes, such as those coding for chlorophyll a/b-binding (Cab) proteins or ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). They therefore provide a convenient system expressing recombinant without background interference. Transfection protoplasts with vectors bearing the Lhcbl*1 Cab gene under control 35S promoter cauliflower mosaic virus (CaMV) resulted in appearance significant amounts specific transcripts, but corresponding polypeptides, inferred northern and western blot analysis, respectively. The use an expression vector carrying translational enhancer Ω tobacco (TMV) strongly enhanced transfected products: analysis clearly revealed Lemna gibba Lhcb2*1, tomato Lhcb2*1 psaD, pea rbcS products. Molecular weight estimations newly synthesized polypeptides indicated that each was promptly processed into its mature-cleaved form within protoplasts. This occurred despite lack absence thylakoid network.
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