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[Molecular cloning and expression analysis of sucrose synthase gene from Dendrobium officinale]
Hengling Meng1, Chengli Duan, Fenghui Xiao
1Institute of Chinese Medical Materials, Yunnan Agricultural University/Yunnan Provincial Good Agricultural Practice Center of Chinese Medical Materials, Kunming 650201, China. menghengl@163.com
Objective:
Clone of sucrose synthase of Dendribium officinale and expression analysis, to provide the theory basis for research the relationship between polysaccharide synthesis of D. officinale and sucrose synthase activity.
Method:
According to homologous sequence of sucrose synthase gene on GenBank, application the technology of RT-PCR and RACE, clone the full length of D. officinale. Target gene amplified with T vector was transformed into competent E. coli. BL21, IPTG induced expression, SDS-PAGE analysis.
Result:
A full length cDNA encoding sucrose synthase was isolated from the D. officinale, named DOSS1, the GenBank accession number is HQ856835, the cDNA is 2781 bp in length containing an open reading frame of 2424 bp encoding 807 amino acids with a predicted molecular mass of 92.3 x 10(3), the deduced amino acid sequence of D. officinale sucrose synthase shares 95% identity with Mokara yellow (AF530568); shares 90% identity with Oncidium goldiana (AF530567); shares more than 80% with other monocotyledonous plants.
Conclusion:
Cloned the sucrose synthase gene and induced an obvious band successfully.