Premium
pTRIDENT, a novel vector family for tricistronic gene expression in mammalian cells
Author(s) -
Fussenegger Martin,
Mazur Xenia,
Bailey James E.
Publication year - 1998
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19980105)57:1<1::aid-bit1>3.0.co;2-m
Subject(s) - cistron , biology , gene , cloning (programming) , multiple cloning site , genetics , enhancer , vector (molecular biology) , transcription (linguistics) , microbiology and biotechnology , gene expression , rna , recombinant dna , linguistics , philosophy , computer science , programming language
We constructed tricistronic expression vectors for the simultaneous and coordinated expression of three independent genes in mammalian cells. One single promoter allows high level and, in some vectors, adjustable transcription of all three cistrons. Whereas the first cistron is translated in a cap‐dependent manner, the subsequent ones utilize intercistronic regions of viral origin such as the internal ribosomal entry site of poliovirus or the cap‐independent translation enhancer of encephalomyocarditis virus for enhanced translation. Three multiple cloning sites with a total of up to 18 unique restriction sites allow sequential cloning of the genes of interest. The modular structure of this pBluescript®‐based high copy number vector system allows straightforward movement of individual cistrons among members of the pTRIDENT family, and facilitates their combination with existing expression vectors. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 1–10, 1998.