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Characterization of an Ethanol‐Inducible Promoter System in Catharanthus roseus Hairy Roots
Author(s) -
Peebles Christie A. M.,
Gibson Susan I.,
Shanks Jacqueline V.,
San KaYiu
Publication year - 2007
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp070154j
Subject(s) - catharanthus roseus , promoter , transgene , ethanol , reporter gene , gene , gene expression , biology , indole test , biochemistry , chemistry
Efforts to engineer Catharanthus roseus hairy roots to produce commercially significant amounts of valuable compounds, such as the terpenoid indole alkaloids vinblastine and vincristine, require the development of tools to study the effects of overexpressing key metabolic and regulatory genes. The use of inducible promoters allows researchers to control the timing and level of expression of genes of interest. In addition, use of inducible promoters allows researchers to use a single transgenic line as both the control and experimental line, minimizing the problems associated with clonal variation. We have previously characterized the use of a glucocorticoid‐inducible promoter system to study the effects of gene overexpression within the terpenoid indole alkaloid pathway on metabolite production. Here the feasibility of using an ethanol‐inducible promoter within C. roseus hairy roots is reported. This ethanol‐inducible promoter is highly sensitive to ethanol concentration with a concentration of 0.005% ethanol causing a 6‐fold increase in CAT reporter activity after 24 h of induction. The ethanol‐inducible CAT activity increased 24‐fold over a 72‐h induction period with 0.5% ethanol.