Retroviral Delivery of Simian Virus 40 Large T Antigen Gene into Primary Cultured Ovary Cells of the Penaeid Shrimp, Penaeus chinensis: Indirect Evidence of Retroviral Integration

摘要

A simian virus 40 large T antigen (SV40 T) gene-carrying pantropic retroviral vector was constructed, in which the SV40 T gene is expressed from Moloney murine leukemia virus (MoMLV) promoter. Using this gene delivery system, we achieved the transfer and expression of SV40 T gene in primary cultured ovary cells of the peneaid shrimp, Penaeus chinensis. The presence of transferred SV40 T gene and expression of the gene mRNA and protein product in transduced cells were determined by polymerase chain reaction (PCR), reverse transcription-polymerase chain reaction (RT-PCR), and immunofluorescence staining, respectively. Although the SV40 T gene failed to induce transformation of shrimp ovary cells maintained in MPS medium, the pantropic retroviral gene delivery system described here is a feasible tool for introducing oncogenes into shrimp cells.