Gene expression of GFP in transgenic rice suspension cell cultures.

<p><b>A,</b> Detection of GFP mRNA in non-transformed (NT) and selected transgenic rice suspension-cultured cells, including αAmy3sp-2, -5, and -9; CIN1sp-3, -5, and -9; and 33KDsp-1, -5, and -9. Total RNAs were isolated from NT and transgenic rice suspension-cultured cells, and then subjected to RT-PCR with specific <i>GFP</i> primers. The rice <i>Act1</i> mRNA was used as an internal control. <b>B,</b> The proportion of medium GFP protein in various transgenic suspension cell lines. Total soluble proteins were extracted from each cell line, with a cell volume of 0.5 mL, in 1 mL of extraction buffer. A total of 1 mL of culture medium for each line was collected. Twenty microliters of total cellular protein (C) and 20 μL of total extracellular protein (M) were subjected to Western blot analysis using GFP antibodies. GFP proteins within cells and medium were detected, and compared with the total amount of GFP protein. The proportion of medium GFP for each independent cell line is presented as the total GFP in medium compared with the total GFP. A dark gray bar indicates medium GFP, while a light gray bar indicates cellular GFP. Error bars indicate the standard deviation of three Western blot replicates from two biological repeats.