Identification And Detection Of Components Of Duck Origin In Foodstuff And Feedstuff Using PCR-mtDNA

At present, the identification of components of animal origin has been involved in the food and meat industry, trade, market and the restaurant industry, and other fields. In view of the demands to identify and detect of components of poultry origin, the experiment is designed to clone COⅢand to identify components of duck origin in foodstuff and feedstuff.Using the duck muscle DNA as template, the mitochondrial cytochrome oxidase III(COⅢ) of duck were amplified by PCR-mtDNA(Genebank accession: DQ655706 see appendix 3).To analyze the cloned sequence, it contains 784bp nucleotides of COⅢgene and partial ATPase6 and tRNA-Gly gene. Homology analysis indicated that the cytochrome oxidase III(COⅢ)is relatively conservative, so molecular evolution can be analyzed with difference of homology. The homology of nucleotide sequences and amino acid sequence results that it is nearly consistent of structure, there are only some changes in coefficient of distance among species.Cloning and analyzing of duck mt-DNA COⅢlay a foundation for identifying and detecting components of duck origin in foodstuff and feedstuff using PCR mtDNA. The sequence of mtDNA can be analyzed with BLAST software and Bioedit software, and also special primer for identification and detection components of duck origin in foodstuff and feedstuff can be designed using PCR mtDNA because of the special character of COⅢ. Using the muscle DNA of various animal and feedstuff that contains components of duck origin as template to amplify, through repeating experiment then the primer that was confirmed is very special and much stable. The primer (P3:5′-CTCATCTATCCTGCTAGCCGC-3′,P4:5′-TACCAACCCCCGAAC TA GGC-3′)can only amply sequence of duck but not can amply sequence of any other animals. The special sequence contains 226bp by PCR,the amplified product was cloned,sequenced and analyzed .The result showed 100% homology with duck mtDNA COⅢgene, which proved the accuracy of special primer. The research that used different consistence of DNA by PCR was to do sensitive experiment with P3 and P4.The result showed that the primer has much speciality and rather sensitivity. So it is a way to identify and detect of duck origin in foodstuff and feedstuff.