Hybrid Gateway Binary Vector System for Cloning of Promoter and cDNA -Application for Promoter Analysis-

Abstract

Previously, we constructed Dual Site Gateway Binary Vector for cloning two genes on separate position. In this paper we report the new Gateway Binary Vector (R4 pGWB) to connect two DNA fragment using a part of Multisite Gateway system. R4 pGWB has attR4-CmR-ccdB-attR2 acceptor site and by the LR reaction using L4-promoter-R1, L1-cDNA-L2 and R4 pGWB, we can make promoter::cDNA construct easily. We also made R4 pGWB series for tag/reporter fusion at C-terminal of cDNA. These new vectors were named Hybrid Gateway Binary Vector system and could be a powerful tool for promoter swapping experiments.
In the report we show the promoter analysis of the gene that expressed in guard cell, as example of application of this system.