Construction Of Plant Expression Vector Of T35s Promoter And VP₄-ST Expression In Alfalfa

It reported that direct duplication of CAMV35S promoter could enhance heterologious gene expression. According to this theory , this research enzymed D35S (double 35S) promoter from the plasmid pBLG and linearized to plasmid vector pUC18. The sequence showed that the D35S was inserted into pUC18 successfully. T35S which contains two A District and four B District was constructed by D35S direct duplication with D35S of vector pJIT60. The promoter of T35S was linearized to plasmid vector pUC18 and sequenced. The promoter of T35S was linearized with pBLGvp4-st. Plant expresses vector pBLGT35svp4-st was constructed by enzyming, linearizing and translating.Alfalfa contained the plant expression vector pBLGT35svp4-st and pBLGvp4- st by Agrobacterium-mediated transformation and antibiotics( Kan) many filtrate.Result: The sequence analysis showed the homoeology of tow A District in T35S promoter are 83.50﹪, homoeology of four B District are 80.31﹪. Plant express vector pBLGT35svp4-st was enzymed by SacI and BamHI. The results showed that the full length of fragment is 1491 bp. The protein of transformed plant was tested by western-Dot and ELISA. It showed significant hybridized signal.