The improvement of amorpha-4,11-diene production by a yeast-conform variant.
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.
Kong
J-Q
JQ
Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, PR China. jianqiangk@imm.ac.cn
Jiang-Qiang
K
K
Wang
W
W
Wei
W
W
Wang
L-N
LN
Li-Na
W
W
Zheng
X-D
XD
Xiao-Dong
Z
Z
Cheng
K-D
KD
Ke-Di
C
C
Zhu
P
P
Ping
Z
Z
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
D000081223:Polycyclic Sesquiterpenes; D012717:Sesquiterpenes; C515348:amorpha-4,11-diene; D019883:Alkyl and Aryl Transferases; C403714:amorpha-4,11-diene synthase