Cloning And Identifing Stress-inducible Promoter Rd29A Of Zoysia Japonica Steud

The promoter Rd29A is induced by drought, high-salt and cold. It is also a ideal inducible promoter in gene engineering of plant resist contrary. Our purpose is to isolate Rd29A promoter gene fragment from Zoysia japonica Steud genomic DNA. Then measuring, comparing and analyzing the order, and indentifying the function of the promoter Rd29A. We screen out inducible promoter to suit and promote transfer-gene technology in turfgrass. The study will be very important for turfgrass growth in resist contrary.According to the reported sequence of promoter Rd29A, designe and synthesize distinctive primers. We amplify the DNA fragments of Rd29A promoter through method of PCR from Zoysia japonica Steud genomic DNA, then measuring, comparing and analyzing the order. It indicates DNA sequence have 41.65% homology with the Rd29A promoter which has been submitted to GenBank. The accession number is EU346948 in GenBank.We utilize the gene of green fluorescence protein (GFP) andβ-GUS coronidaSe (GUS) as reporting genes, four transient plant expression vectors pBIG(35S-GFP), pBIRG(Rd29A-GFP) and pBI(35S- GUS), pBIR(Rd29A-GUS) are constructed. They are transformed the epidermis cells of onion by particle bombardment. After coerce inducib, detecting purpose fragment to regulate and control gene expression in acceptor cell, identify whose function thereby.Result shows that cloning purpose fragment has the function of promoter, and it can regulate lower reaches gene expression intensely in drought, high-salt and cold.