Construction of Yeast-one-hybrid Bait Vector and Screening of Interacted Proteins of HbCoi1

Key words: 
• HbCoi1 /
• pHIS vector /
• auto-activation /
• yeast-one-hybrid

Abstract: Jasmonic acid（JA） is one of the most important phytohormones for regulation of latex biosynthesis in Hevea brasiliensis. HbCoi1 is the receptor of JA signaling pathway. Analysis of the gene expression of HbCoi1 is of great significance to reveal molecular mechanism of latex biosynthesis in H. brasiliensis. In this context,the promoter of HbCoi1 was PCR amplified and inserted into pHis2. 1 vector to construct bait vector p HIS-HbCoi1 for yeast-one-hybrid. The vector was then transformed into Y187 yeast strain and cultured on the SD medium to test the auto-activity of the transcription. The growth of the strain containing bait vector was inhibited on the SD medium added with 10 mmol·L^-1 3-AT. Furthermore,p HIS-HbCoi1 vector and p GADT7-rec2 cDNA library of H.brasiliensis were co-transformed into Y187 strain,and several proteins interacted with HbCoi1 promoter were identified,including DNA binding protein,ribosomal proteins and uncharacterized proteins,which lays a foundation for further study of gene expression of HbCoi1.