An Update of pTRIDENT multicistronic expression vectors: pTRIDENTs containing novel streptogramin-responsive promoters

摘要

We present an update on the pTRIDENT multicistronic mammalian expression vectors and their implications in various metabolic engineering and therapeutic applications. The pTRIDENT vector family has been expanded by construction of a new set of pTRIDENT-based vectors containing constitutive promoters of human origin (ubiquitin C and EF-1#alpha# promoters) and selectable markers (zeocin resistance) and expressing different reporter genes (secreted alkaline phosphatase (SEAP) and the secreted single-chain urokinase-type plasminogen activator (low-M-r u-PA)). In addition, we have constructed pTRIDENT derivatives with novel streptogramin-repressible and strep-togramin-inducible promoters for simultaneous and adjustable expression of three different transgenes. Streptogramin-inducible and teracycline-repressible pTRIDENT derivatives were used to simultaneously control expression of three fluorescent proteins in mammalian cells: the enhanced cyan fluorescent protein (CFP), and the enhanced yellow fluorescent protein (YFP). Owing to their modular structure, the pTRIDENT vector family represents a construction kit for the design of novel multicistronic expression constructs.