title: The improvement of amorpha-4,11-diene production by a yeast-conform variant.
abstract: OBJECTIVE
  To investigate the effect of the yeast-conform variant of the Artemisia annua gene encoding for amorpha-4,11-diene synthase (ADS) on the production of amorpha-4,11-diene in a transformed yeast.
  
  
  RESULTS
  The ADS gene was mutated to the yeast-conform variant ADSm. The ADSm synthesis was performed based on step-by-step extension of a short region of the gene through a series of polymerase chain reactions (PCR). The artificial ADSm gene contained codons preferred by the yeast translation machinery. The sequence was then integrated into a yeast expression vector pYeDP60. The fusion construct was active and the transformed yeast cells produced higher level of amorpha-4,11-diene compared with the plant gene-transformed yeast cells.
  
  
  CONCLUSIONS
  Strains transformed with the yeast-conform allele (ADSm) were more efficient in terms of production of amorpha-4,11-diene than those transformed with the plant gene.
  
  
  CONCLUSIONS
  We demonstrated that yeast-conform allele of foreign genes by serial PCR reactions can be a solution to low efficiency of heterologous gene expression in Saccharomyces cerevisiae cells.
authors.affiliation: Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, PR China. jianqiangk@imm.ac.cn
authors.forename: J-Q
authors.initials: JQ
authors.lastname: Kong
authors.forename: K
authors.initials: K
authors.lastname: Jiang-Qiang
authors.forename: W
authors.initials: W
authors.lastname: Wang
authors.forename: W
authors.initials: W
authors.lastname: Wei
authors.forename: L-N
authors.initials: LN
authors.lastname: Wang
authors.forename: W
authors.initials: W
authors.lastname: Li-Na
authors.forename: X-D
authors.initials: XD
authors.lastname: Zheng
authors.forename: Z
authors.initials: Z
authors.lastname: Xiao-Dong
authors.forename: K-D
authors.initials: KD
authors.lastname: Cheng
authors.forename: C
authors.initials: C
authors.lastname: Ke-Di
authors.forename: P
authors.initials: P
authors.lastname: Zhu
authors.forename: Z
authors.initials: Z
authors.lastname: Ping
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism*; D031172:Artemisia annua / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D005818:Genetic Engineering / Q000379:methods*; D016681:Genome, Fungal / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D000081223:Polycyclic Sesquiterpenes; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D012717:Sesquiterpenes / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: D000081223:Polycyclic Sesquiterpenes; D012717:Sesquiterpenes; C515348:amorpha-4,11-diene; D019883:Alkyl and Aryl Transferases; C403714:amorpha-4,11-diene synthase