Cloning and Expression Analysis of the Promoters of OfLCYB and OfLCYE in Osmanthus fragrans

doi:10.13560/j.cnki.biotech.bull.1985.2017-0706

Abstract

Abstract: Lycopene cyclization is an important branching point in the synthesis of carotenoids. There are two kinds of lycopene cyclase LCYB and LCYE in plants. Previous studies have found that the differential expressions of OfLCYB1,OfLCYE1 and other genes determine the contents of downstream products of carotenoid metabolism in different varieties of Osmanthus fragrans. In this study,1028 bp and 904 bp of the OfLCYB and OfLCYE promoters were cloned by genome walking with the genomic DNA of cultivar ‘Yanhong Gui’ as the template. The bioinformatics analysis discovered two promoter sequences that contained TATA-box and CAAT-box,also the TCA-element of salicylic acid,and some photoresponsive elements such as Sp1,ACE,Box4,and G-box. The OfLCYB promoter also includes abscisic acid response element ABRE,gibberellin response element P-box. OfLCYE promoter includes gibberellin response element GARE-motif and thermal response element HSE. The cloned promoter fragment was recombined with pBI121 vector,and the plant expression vector was constructed and transferred into Agrobacterium tumefaciens. The tobacco leaf was used for transient expression. The results proved that OfLCYB and OfLCYE promoter fragments all had the function of driving downstream reporter gene expression.

Key words: Osmanthus fragrans, promoter, LCYB, LCYE, transient expression

SHEN Zi-you, ZHANG Chao, DONG Bin, FU Jian-xin, HU Shao-qing, ZHAO Hong-bo. Cloning and Expression Analysis of the Promoters of OfLCYB and OfLCYE in Osmanthus fragrans[J]. Biotechnology Bulletin, 2018, 34(1): 137-143.

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