Cloning of a flower-specific expression promoter from Arabidopsis thalianaand its plant expression vector construction
Bing-you Fan; Shui-ping Gao; Xiao-gai Hou; Guo-an Shi
Forest Science and Practice 12(4): 201-205
Given the sequence of Chs gene promoter from Arabidopsis thaliana reported in Gen Bank (AF248988), a pair of specific Pcr primers was designed with the Primer Premier 5.0 software. Pcr products of about 0.5 kb were successfully amplified with the genome Dna of A. thaliana as a Dna template and Taq polymerase as Dna polymerase. The purified Pcr products were ligated to the p MD18-T vector. The sequencing result showed that the Chs promoter from A. thaliana was 531 bp long. Sequence alignment analysis based on the Dnaman software revealed that the sequence similarity between the cloned promoter and target promoter (AF248988) was up to 100%. Online Place analysis indicated that the Chs promoter contained cis-elements such as TATA-box, CAAT-box, pollen-box, G-box, ACGT-containing element, R response element, Myb recognition element and TACPy AT-box. At the same time, a plant expression vector p At Chs::Gus which fused the Chs promoter and the marker gene Gus was successfully constructed.