Abstract:

Genomic DNA was extracted from leaf, and total RNA was isolated from latex of rubber tree. REF promoter fragment of 378 bp and the V-PPase fragment of 2310 bp was cloned by PCR using specific primers. Sequence analysis showed that, the cloned V-PPase gene fragment was identical with that logged on GenBank, and the cloned REF promoter fragment shared high identities with four submitted REF promoter sequences in NCBI at a rate of 98.68 % and 99.47 %, respectively. Then the two fragments was cloned into the plant expression vector pCAMBIA1301, pCAMBIA2301 and pCAMBIA3301 by restriction enzyme digestion and ligation to get the recombinant expression vector pC1301RV, pC2301RV and pC3301RV contained hygromycin phosphotransferase Ⅱ(hpt Ⅱ) gene , neomycin phosphotransferaseⅡ(npt Ⅱ) gene and bialaphos resistance(bar) gene, respectively, which might express V-PPase gene efficiently in laticifer of rubber tree. The expression vectors were transferred into Agrobacterium tumefaciens strain EHA105 by freeze-thaw method. This work set a good foundation for further genetic transformation of rubber tree.