S. Moser et al., An update of pTRIDENT multicistronic expression vectors: pTRIDENTs containing novel streptogramin-responsive promoters, BIOTECH PR, 16(5), 2000, pp. 724-735
We present an update on the pTRIDENT multicistronic mammalian expression ve
ctors and their implications in various metabolic engineering and therapeut
ic applications. The pTRIDENT vector family has been expanded by constructi
on of a new set of pTRPDENT-based vectors containing constitutive promoters
of human origin (ubiquitin C and EF-1 alpha promoters) and selectable mark
ers (zeocin resistance) and expressing different reporter genes (secreted a
lkaline phosphatase (SEAP) and the secreted single-chain urokinase-type pla
sminogen activator (low-M-r u-PA)). In addition, we have constructed pTRIDE
NT derivatives with novel streptogramin-repressible and streptogramin-induc
ible promoters for simultaneous and adjustable expression of three differen
t transgenes. Streptogramin-inducible and tetracycline-repressible pTRIDENT
derivatives were used to simultaneously control expression of three fluore
scent proteins in mammalian cells: the enhanced cyan fluorescent protein (C
FP), the recently isolated red fluorescent protein (RFP, also designated ds
Red), and the enhanced yellow fluorescent protein (YFP). Owing to their mod
ular structure, the pTRIDENT vector family represents a construction kit fo
r the design of novel multicistronic expression constructs.