Identification of rodent residues by DNA barcoding technology in plague areas / 中华地方病学杂志

ABSTRACT

Objective To identify rodent residues using DNA barcoding technology in plague areas of Shaanxi and to analyze the feasibility of DNA barcoding method.Methods DNA extraction,PCR,electrophoresis and sequence determination and alignment were used to determine the cytochrome C oxidase Ⅰ (COI) gene sequence from 62 residues of 7 species identified by morphology in 8 different parts.COI gene sequence was analyzed using BLAST software of American National Center for Biotechnology Information (NCBI) for sequence homology comparison and the phylogenetic tree was constructed by using the neighbor joining(NJ)method of COI gene sequence.Results In addition to the hair,CO I genes of the feet,the tail,the fur,the muscle,the ribs,the ear,and the eye were amplified,respectively,and the size of amplified fragment was similar to the size of the product with an expected fragment(700 bp),and the fragment was a single band.COI genes of 59 residue specimen were obtained by DNA sequencing and there were high degree of homologies between CO I gene sequences of Meriones unguiculatus,Cricetulus barabensis,Meriones meridianus,Dipus sagitta,Phodopus roborovskii,Rattus norvegicus and Allactaga sibirica and their corresponding host genes in NCBI(99.0％,98.1％,99.8％,98.9％,99.5％,99.1％,98.3％).NJ method showed that 7 NJ phylogenetic trees were constructed with COI sequence of 59 species residues.The CO I sequences of same rodent with different residues were clustered into one group.Condusion DNA barcoding technology can identify host animal residues in plague areas,and the identification results are reliable.