Gateway Binary Vectors for Expression of Tag Fused Protein in Plant Cell

Abstract

We made Gateway Binary Vectors to facilitate the cloning in binary vector. Especially, we were interested in and made tag-adding Gateway Binary Vectors. Previously, we reported GFP, 6xHis, FLAG, 3xHA, 4xMyc, 10xMyc and GST adding vectors. Recently, we made 3 type of T7 adding vectors. They are (no promoter, C'-T7), (35S promoter, C'-T7) and (35S promoter, N'-T7).
To check the expression and tag fusion in plants, we made 35S pro::tag::GUS and 35S pro::GUS::tag constructs using GUS entry clone and each Gateway Binary Vectors. They were introduced into tabocco BY-2 cells and analyzed by immunoblotting using anti-tag antibody and anti-GUS antibody. We also mesured GUS activity and estimated specific activity of tag fused GUS protein.