TY - JOUR
T1 - A plasmid family containing two different expression cassettes suitable for immunomodulation and genetic immunization
AU - Ciafrè, Silvia A.
AU - Rinaldi, Monica
AU - Vespignani, Isabella
AU - Parrella, Paola
AU - Seripa, Davide
AU - Signori, Emanuela
AU - Ria, Francesco
AU - Farace, Maria Giulia
AU - Fazio, Vito M.
PY - 1998/7
Y1 - 1998/7
N2 - 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 the other. We recommend the use of pRC110 for genetic immunization and immunological response studies, when the concomitant local production of an immunogenic peptide and of a syngeneic immunomodulating cytokine is required.
AB - 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 the other. We recommend the use of pRC110 for genetic immunization and immunological response studies, when the concomitant local production of an immunogenic peptide and of a syngeneic immunomodulating cytokine is required.
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U2 - 10.1006/plas.1998.1339
DO - 10.1006/plas.1998.1339
M3 - Article
C2 - 9657937
AN - SCOPUS:0031827185
VL - 40
SP - 84
EP - 89
JO - Plasmid
JF - Plasmid
SN - 0147-619X
IS - 1
ER -