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.
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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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Kai, G., Miao, Z., Zhang, L. et al. Molecular cloning and expression analyses of a new gene encoding 3-hydroxy-3-methylglutaryl-CoA synthase from Taxus × media . Biol Plant 50, 359–366 (2006). https://doi.org/10.1007/s10535-006-0050-0
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DOI: https://doi.org/10.1007/s10535-006-0050-0