A plasmid family containing two different expression cassettes suitable for immunomodulation and genetic immunization

Plasmid
S A CiafrèV M Fazio

Abstract

We have developed an improved eukaryotic expression vector that consists of two distinct, complete, and differentially regulated transcription units. The peculiarities of this prototype vector, named pRC110, are represented by two different strong promoter/enhancer sequences, cytomegalovirus and Rous sarcoma virus, that independently drive transcription of two recombinant cDNAs, which may be easily cloned into specific rare restriction sites. Moreover, we describe a simple way to introduce an optimal translational start site context 5' to any peptide to be cloned in our vectors, thus allowing the correct and efficient expression of even a single part of a larger gene or a short synthetic peptide lacking its own AUG and neighboring regions. We demonstrate the in vivo expression efficacy of pRC110 for use in genetic vaccination through direct intramuscular gene transfer: specific antibodies are raised against one of the encoded peptides 3 weeks after muscle injection, and efficient transcription of the other syngeneic cDNA, mouse interleukin-2, is shown. The development of a "family" of vectors directly deriving from pRC110 is also described, with the common property that one of the encoded proteins may modulate the effects of th...Continue Reading

References

Jan 4, 1996·The New England Journal of Medicine·W M McDonnell, F K Askari
Jun 1, 1995·Immunological Reviews·F K StevensonR E Hawkins
Aug 1, 1995·Immunity·D M Pardoll, A M Beckerleg

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Citations

Oct 6, 2007·Expert Opinion on Biological Therapy·Pieranna ChiarellaVito Michele Fazio
Aug 23, 2006·FEMS Microbiology Letters·Manuela PardiniAntonio Cassone

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