Characterization of cell lines that inducibly express the adeno-associated virus Rep proteins.

摘要: The replication (rep) gene of adeno-associated virus (AAV) is involved in AAV DNA replication, regulation, and inhibition cellular transformation induced by various oncogenes. To study the rep gene's antiproliferative effects, we have developed cell lines which express proteins under control an inducible mouse metallothionein transcription promoter. Rep78 protein produced these binds to terminal repeat sequences vitro supports trans activation p40 promoter vivo. These are capable assembling infectious viruses containing a mutant or vector bearing heterologous gene. Growth rate colony formation efficiency assays indicated that expression substantially altered proliferation. Long-term induction followed removal inducing agent suggested constitutive Rep does not necessarily result death cells can recover from cytostatic effects. Flow cytometry analysis presence increased population S phase cycle. Thus effects may be realized interference with replication.