Analysis and stability of the Respiratory Syncytial Virus antigen in a T3 generation of transgenic tomato plants

The integration, expression, and stability of the Respiratory Syncytial Virus (RSV)-F protein was analyzed in a T-3 generation of transgenic cherry tomato, Solanum lycopersicum L. cv. Swifty Belle, plants. Expression of the RSV-F antigen, under the control of the fruit-specific promoter E-8, was investigated in T-3 plants derived from a transgenic line, identified as #120. Transgene integration of the RSV-F gene in the T-3 generation was initially determined by polymerase chain reaction (PCR). PCR analysis from line 120-7-2 revealed that all T-3 plants were homozygous for the transgene; whereas, line 120-6-4 showed segregation for the transgene. Enzyme-linked immunosorbent assay (ELISA) was used to quantify levels of RSV-F protein in these plants, and protein levels ranged from 0-22 mu g/g of fresh weight, with an average of similar to 3 mu g/g fresh weight. Southern blot analysis of the highest expressing plants revealed presence of a single copy of the RSV-F transgene in these plants.