| In mammal, SRY/Sry is a sex determining gene putatively. All relative researches have shown that the presence of SRY/Sry gene leads to male phenotype, or else, female phenotype forms. The sex determination is started by gene SRY/Sry, which happens between 10.5d.p.c. and 12.5 d.p.c, and SRY/Sry synchronously presents at this time.In our previous experiment, male pheynotype of XY genotype individual was successfully developed into female phenotype mice (sex reversal mice) by using nucleic acid vaccine that might block the pathway of Sry during embryo sex determination. There are still some more problems must be resolved about the vaccine in further research, such as optimizing the way of injection and studying the mechanism of the vaccine. In our research,The aims of present investigations would focus on: 1. verifying the validity of the vaccine. 2. studying the correlativity between the inject method and immune efficiency by comparing the immune efficiency of the two methods.Two immune methods, intramuscular injection and gene gun injection, were selected. At the same time, we improve the method of vaccine preparation and young mice detection. For example, we remove the endotoxin of the vaccine by reagent kit and detect the young mice dual by nest PCR and southern blot technique.Results: For intramuscular injection, there are two sex reversal mice in 181 young mice, the immune efficiency is about 1.1%. For gene gun injection, there are two sex XY female reversal mice in 240 young mice, the immune efficiency is about 0.8%.Although the results did not distinguish the two immune methods, those indicated that there might be some other factors instead of immune methods play more important role in present gene vaccine administration. It was clearly shown that Sry gene vaccine was validity (but with low efficiency) by blocking male phenotype development during embryo for getting sex reversal mice. Those results were verified by both nest PCR and southern blot technique at same the time. |
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