A polyomavirus enhancer mutant confers ubiquitous high transcriptional efficiency to the SV40 late promoter.
To identify expression plasmids with high efficiency of transcription and with a broad tissue and cell range, we have constructed a recombinant vector combining the late SV40 promoter and the polyomavirus regulatory region derived from a mutant (PyNB11/1) which displays a very wide host range. We show that these recombinant enhancer-promoters are efficient drivers for heterologous gene transcription and expression in vitro in all mouse and human cells tested. The most active combination we identified contained the mutant enhancer (PyNB11/1) in the late orientation. This construct was able to promote a high efficiency of expression without significant fluctuation between cells of different tissutal origin or different differentiative stage. A possible interpretation of these results is discussed.
Istituto Pasteur Fondazione Cenci-Bolognetti, Dipartimento di Biopatologia Umana, Università di Roma La Sapienza, Italy.
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D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D002522:Chlorocebus aethiops; D004742:Enhancer Elements, Genetic; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D008113:Liver Neoplasms; D051379:Mice; D008969:Molecular Sequence Data; D011120:Polyomavirus / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D013539:Simian virus 40 / Q000235:genetics*; D013724:Teratoma; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
D015500:Chloramphenicol O-Acetyltransferase