Figure 2

Transient expression of mBFP and mBFP-mS ₁ C fusion protein in tobacco leaves by agroinfiltration. A. The mBFPvvD8 (D8) or mBFPvvD9 (D9) gene expression constructs driven by RbcS promoter were transiently expressed in tobacco leaves by agroinfiltration, respectively. The infiltrated tissues were imaged under fluorescence microscope with UV excitation (UV) or bright light (BL). C, Agrobacteria only as control. The magnification is 400 X and the ruler is 20 μm. B. The mBFP or mBFP-mS₁C fusion gene expression cassettes driven by RbcS promoter were transiently expressed in tobacco leaves by agroinfiltration, respectively, and the proteins were targeted to cytosol (Cy, pBin-R-Cy-mBFP), apoplast (Se, pBin-R-Se-mBFP), ER (mBFP, pBin-R-Er-mBFP; mBFP-mS₁C, pBin-R-Er-mBFP-mS₁C), chloroplast (Cp, pBin-R-Cp-mBFP) or mitochondria (Mt, pBin-R-Mt-mBFP), respectively. The infiltrated tissues were imaged under fluorescence microscope with UV excitation (UV) or bright light (BL). C, Agrobacteria only as control. The magnification is 200 X and the ruler is 40 μm. C. The protoplasts were isolated from tobacco leaves that transiently expressed pBin-R-Er-mBFP (mBFP) or pBin-R-Er-mBFP-mS₁C (mBFP-mS₁C) constructs, and were imaged by confocal microscope. BL, bright light; Red, chlorophyll auto-fluorescence; Blue, mBFP blue fluorescence. The magnification is 1,000 X and the ruler is 20 μm. D. RT-PCR assay was used to measure the relative abundance of mBFP mRNA in tobacco leaves which transiently expressed pBin-R-Er-mBFP (Lanes 1, 3 and 5) or pBin-R-Er-mBFP-mS₁C (Lanes 2, 4 and 6) constructs. The Nt EF-1α was used as a control. Three independent experiments were carried out. M, 100 bp marker. E. The relative abundance of mBFP mRNA to that of Nt EF-1α was quantitated by Image J. mBFP, pBin-R-Er-mBFP construct; mBFP-mS₁C, pBin-R-Er-mBFP-mS₁C construct.