| Objective To construct binary vectors which contains HPV16 E6 and E7 gene for edible vaccine in transgenic tomato. The toxic effect of transgenic soybean feed on reproductive system was evaluated in male mice.Methods E6 and E7 sequences were analyzed with Vector NTI8.0 software for codon-optimization. The molecular adjuvant gene LTB and regulatory sequences, such as KOZAK and SEKDEL endoplasmic reticulum retention motif were combined to fused genes. These genes were synthesized with overlapping PCR, and cloned into T vector, confirmed with PCR and DNA sequencing. The genes were introduced into pX6-GFP for tomato transformation.After feeding male mice with transgenic soybean containing transgenic genes, 5-enolpyruvyl shikimate-3-phosphate synthase (EPSPS), Nopaline Synthase (NOS) Terminator and Cauliflower mosaic virus 35S promoter(CaMV35S) for 120 days, we assess the potential transfer of diet-derived DNA to animal tissues after consumption of GM plants with PCR. Pathological changes of testis tissue was examined with Histologic microtomy. The number of sperm at different metosis phase were analyzed with flow cytometry. The integrity of chromatin of sperm were tested by comet assay.Results E6 and E7 gene of HPV 16 were successfully optimized and synthesized, then constructed to the binary vectors for tomato transformation. Foreign genes can't detected by PCR in testis tissue of mice fed with transgenic-soybean. Microscopy examination showed no pathological changes in testis.The results showed no significant difference in numbers of sperm at different metosis phase and chromatin integrity between animals raised with the transgenic feed and animals raised with the conventional feed. Conclusions Optimized E6 and E7 genes were synthesized with overlapping PCR and constructed to the binary vectors for edible vaccine in tomato. Testes of male mice as target organs, no toxicological effects of transgenic feeding on reproductive system was showed...
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