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ACTA HORTICULTURAE SINICA ›› 2014, Vol. 41 ›› Issue (6): 1236-1244.

• Research Notes • Previous Articles     Next Articles

Cloning and Expression Analysis of Deoxyoxylulose-5-phosphate Synthase Gene Related to Aroma from Jasminum sambac and Isolation of Its Promoter

SUN Jun1,CHEN Gui-xin1,*,YE Nai-xing1,2,*,Lü Shi-heng1,LIU Zhi-qin3,HUANG Wei1,and LIN Zhi-da1   

  1. (1College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou 350002,China;2The Institute of Tea Science and Technology and Economy,Fujian Agriculture and Forestry University,Fuzhou 350002,China;3College of Life Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
  • Received:2013-12-13 Online:2014-06-25 Published:2014-06-25

Abstract: The full length cDNA sequence of Deoxyoxylulose-5-phosphate synthase gene(JsDXS)was cloned from Jasminum sambacby using RT-PCR and rapid amplification of cDNA ends(RACE)method. According to the 2 505 bp sequence,the promoter of JsDXS was isolated using Genome Walking Method. The gene expression pattern in blooming was analyzed by the real-time PCR. Gene structure analysis showed that the full length cDNA contained a 2 145 bp open reading frame(ORF),encoding a protein of 714 amino acid. There were several conserved domains,such as TPP-DXS,TPP-PYR-DXS- TK-like,Transketolase C,etc. One 745 bp sequence was found to be the promoter of JsDXS,which contained many cis-acting elements,such as TATA-box,T/GBOXATPIN2(related to jasmonate signaling),MYCCONSENSUSAT(related to cold stress),CIACADIANLELHC(necessary for circadian expression).The real-time PCR analysis revealed that the gene expression at 18:00 was low and started to increase and reached a maximum at 22:00,and then started to decrease and reached a minimum at 2:00. This phenomenon perhaps illustrated that the expression of JsDXS was regulated by circadian.

Key words: Jasminum sambac, aroma, deoxyoxylulose-5-phosphate synthase, promoter, cis-acting element, expression analysis

CLC Number: