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pTRIDENT, a novel vector family for tricistronic gene expression in mammalian cells

✍ Scribed by Martin Fussenegger; Xenia Mazur; James E. Bailey

Publisher
John Wiley and Sons
Year
1998
Tongue
English
Weight
280 KB
Volume
57
Category
Article
ISSN
0006-3592

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✦ Synopsis

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 pBluescriptா-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.

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