Clone and activity analysis of promoter fragment of the F3'5'H gene of Lycium ruthenicum Marr. and its albino fruits
The promoter fragment of the F3'5'H gene of Lycium ruthenicum Murr. and its al-bino fruit was cloned by 3 times of genome walking by Genome Walking Kit (TaKaRa) and sequenced. The bioinformatics analysis showed that the sequence homology of two promoters was 90. 3%, the sequences were submitted to PlantCare to predict the promoter elements. The results showed that both promoters have TATA-Box, CAAT-Box, TC-rich repeats, WUN-motif, Sp1, Box I, G-box, skin-1 and ARE. But promoter TGACG-motif predicted in Lycium ruthenicum Murr. was not predicted in the albino fruit promoter. With 2 promoters combined with the GUS reporter gene to construct the plant transient expression vector, and transient transformed tobacco by Agrobacterium -mediated Method, histochemical staining was performed to determine the promoter activity. The results showed that both promoters had activities. Then the GUS gene expression level was analyzed by Real-time PCR. The results showed that the GUS gene expression level driven by Lycium rutheni-cum Murr. promoter was 3.09 times of its albino fruits.
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