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Biologia Plantarum

, Volume 50, Issue 3, pp 359–366 | Cite as

Molecular cloning and expression analyses of a new gene encoding 3-hydroxy-3-methylglutaryl-CoA synthase from Taxus × media

  • G. Kai
  • Z. Miao
  • L. Zhang
  • D. Zhao
  • Z. Liao
  • X. Sun
  • L. Zhao
  • K. Tang
Article

Abstract

A new full-length cDNA encoding 3-hydroxy-3-methylglutaryl-CoA synthase (designated as TmHMGS, GenBank Accession No. AY644708), which catalyses the condensation of acetyl CoA and acetoacetyl CoA to form 3-hydroxy-3-methylglutaryl-CoA as an early step in the taxol biosynthetic pathway, was isolated from young leaves of Taxus × media by rapid amplification of cDNA ends (RACE) for the first time. The full-length cDNA of TmHMGS contained a 1431 bp open reading frame (ORF) encoding a deduced protein of 476 amino acid residues. The deduced protein had an isoelectric point of 5.23 and a calculated molecular mass of about 53 kDa. Amino acid sequence comparison analysis showed that TmHMGS had high similarity with a number of HMGSs ranging from Schizosaccharomyces pombe to humans, with much higher identity with other HMGSs from plants than those from yeast and humans. Phylogenic analysis showed that TmHMGS had closest relationship with HMGS from Pinus sylvestris. Tissue expression pattern analysis showed that TmHMGS expressed in needles and stems at similar level, but no expression could be detected in roots. Expression of TmHMGS was all induced by under different elicitors such as silver nitrate, ammonium ceric sulphate and methyl jasmonate, revealed that TmHMGS was an elicitor-responsive gene.

Additional key words

elicitor expression profile phylogenetic tree analysis TmHMGS yew 

Abbreviations

ACS

ammonium ceric sulphate

DXP

1-deoxyxylulose 5-phosphate

MJ

methyl jasmonate

MVA

mevalonate

ORF

open reading frame

PCR

polymerase chain reaction

pI

isoelectric point

RACE

rapid amplification of cDNA ends

RT-PCR

reverse transcriptase-polymerase chain reaction

SN

silver nitrate

TmHMGS

T. media 3-hydroxy-3-methylglutaryl-CoA synthase

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Copyright information

© Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Praha 2006

Authors and Affiliations

  • G. Kai
    • 1
    • 2
  • Z. Miao
    • 1
  • L. Zhang
    • 3
  • D. Zhao
    • 3
  • Z. Liao
    • 3
  • X. Sun
    • 3
  • L. Zhao
    • 1
  • K. Tang
    • 1
    • 3
  1. 1.School of Agriculture and BiologyShanghai Jiaotong UniversityShanghaiP.R. China
  2. 2.College of Life and Environment ScienceShanghai Normal UniversityShanghaiP.R. China
  3. 3.School of Life SciencesFudan UniversityShanghaiP.R. China

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