Construction and Self-activation Detection of Bait Vector for Yeast Two-hybrid System of TA gene in Acidovorax citrulli

Objective This project aims to construct a bait vector for yeast two-hybrid system of TA gene in Acidovorax citrulli in order to lay the foundation for exploring the gene function involved in the pathogenic mechanism.MethodThe TA gene was amplified by PCR and cloned into the vector pGBKT7.The verified pGBKT7-TA was then transformed into the yeast strain AH109.The activities of report genes ADE2,HIS3 and LacZ were used to detect the self activation of pGBKT7-TA,and the toxicity of pGBKT7-TA was investigated in nutrient deficient medium.ResultThe constructed bait vector pGBKT7-TA had neither self-activation nor toxic effect in the yeast strain AH109.ConclusionThe constructed bait vector can be used for the follow-up work of the yeast two hybrid system,which has laid a foundation for further screening of proteins interacting with TA from the cDNA library of cucumber cotyledons infected by Acidovorax citrulli.