Cloning of Lycopene β-Cyclase Gene and Activity Analysis of its Promoter from Dunaliella bardawil
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Abstract:
The unicellular green alga Dunaliella bardawil has been successfully used for natural production of β-carotene. Lycopene β-cyclase (LycB) is one of the key enzymes for the synthesis of β-carotene, which can catalysis lycopene to form β-carotene. In this research, based on the full-length sequence of LycB cDNA of D. bardawil previously coloned by our group, primers were designed and genomic sequence of LycB were isolated by analysis of the conserved region of LycB gene from other species in database and by using the approach of PCR with genome walking techniques. Sequence results showed that nearly full-length of LycB genomic sequence of 5863 bp was achieved, which included 11 exons and 10 introns. Then, two sets of primers were designed according to the LycB genomic sequence. The promoter and terminator of LycB were obtained using genome walking techniques. Based on the analysis of PlantPAN, some cis-acting elements such as light-induced responsive element and salt- induced responsive element were predicted in promoter sequence, implying that the LycB gene expression may be regulated by light and salt.
