Studies On PCR-SSCP And PCR-RF-SSCP In A Fragment Of OVR Gene

A chick 424 fragment including a restricted ovulator site of occyte vitellogenesis receptor(OVR) gene was amplified using primers designed according to sequence published, PCR conditions were determined by orthogonal designed and PCR product was detected by the PCR-SSCP and PCR-RF-SSCP techniques in Hyline Brown, Silkies,Abor Acres Broiler (AA). The results are as follows:1. PCR products were verified by HaeIII digestion and sequence analysis;2. Orthogonal design can be used to determine the effect of the PCR parameters;3. No polymorphism by using PCR-SSCP analysis in Hyline Brown(SO), Silkies(44)and Abor Acres Broiler(AA,50) was found, and the reason could be attributed to that the fragment is too long;4. Six genotypes were detected in Silkies ,only three genotypes were detected in Hyline Brown and four genotypes were detected in Abor Acres Broiler with PCR-RF-SSCP after digesting PCR products as two small fragments by Haelll ;5.It was demonstrated that the allele frequency of site A,B respectively among three breeds are significantly different;6.Sequence analysis shows that there are two transition at position 61(C -A) and 164(G-A) respectively between A|Ai and AaA2 individual, and the former resulted in corresponding changes of amino acid (Pro-Thr);As for site B, there is one transition at position 324(C-A) between BjBi and 8283 individual.