Abstract：Type II secretory system is associated with secretion of pathogenic proteins of Ralstonia solanacearum. The gsp gene cluster encoding type II secretory system of R. solanacearum consists of 12 gsp genes. All of gsp genes were cloned successfully by designing optimal PCR reaction system suitable for high content of GC. The 12 gsp genes were ligated into bait and prey vector, respectively. The bait and prey pool of yeast two-hybrid system were constructed. The open reading-frames of 12 gsp genes were verified by sequencing to ensure that gsp genes can be expressed rightly in yeast system, which establish the foundation for research interactions between GSP proteins.
