Detection Of PCR-MtDNA Of Rabbit Components In Food

Food safety issues are gradually aroused people’s attention. In order to truly ensure ediblesafety of ordinary consumers, make sure all kinds of animal derived food quality, it presentsnew challenges for animal meat dish in China component detection. The current source of foodtesting of animals have many problems, to detect the source of the animal ingredients in thework of the need, we study in the DNA CO Ⅲ of clone andPCR-mtDNA technology to detectthe source of the rabbit, and done considerable work.By the rabbit muscle tissue of the DNA as a template, with design synthetic P1, P2as aprimer, we through the PCR-mtDNA technology, the successful cloned the rabbit mitochondrialDNA COⅢ genes. Through the sequence analysis shows, that the rabbit mitochondrial DNAsequence including COⅢ gene sequences of the784bp. The sequence contains a startingpassword son (ATG), a not completely ended anticodon (T) and3’end of the downstreamATPase6and5’ the upstream tRNA-Gly.Rabbit mitochondrial DNA CO Ⅲgene cloning and sequence analysis of the success of theresearch in the PCR-mtDNA as a new technology, to detect the rabbit meat of source sexcomposition laid a theoretical foundation. Through a variety of biological analysis software toanalyze the mitochondrial DNA sequence, and then, according to the rabbit mitochondrial DNACOⅢ gene has the characteristics of the specific to design the specificity of primers P3(22bp),,P4(22bp), And then the PCR-mtDNA method to detect rabbit source sex composition.Using avariety of animals of muscle tissue for PCR amplification DNA template reaction, throughrepeated screening experiment. The last to demonstrate to P1, P2as a primer, can enlarge therabbit DNA fragments, unable to enlarge the DNA of other animals. Explain, primer P1, P2bespecific to the strong and stability good characteristic; To P1, P2for specific primer, through thepolymerase chain reaction (PCR) a rabbit DNA fragments expansion specificity, the segmentsize for230bp. Then will be amplified PCR product to biotechnology company sequenced.Through the analysis and conclusion is already cloning rabbit mitochondrial DNA cytochromeoxidase Ⅲ gene homology is completely consistent, proof of primers, the accuracy of the P1,P2. With10%,2%,1%,0.02%,0.01%,0.002%,0.001%of the content of different gradientDNA template solution PCR amplification, detection specificity primer P3, P4sensitivity. The analysis of experimental results show that specific primer P3, thesensitivity of P4is higher,about0.001%.This paper established the PCR mtDNA technology with meat or feed screening tests ofrabbit source sex composition. This technology has strong specificity, high sensitivity, savestime and effort, detection speed and low cost many characteristics, PCR mtDNA technology canbe used as a test of rabbit meat and feed into conventional method source.