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Species Identification Of Mammals Based On Mitochondrial Cytochrome Oxidase Subunit I Gene Sequencing

Posted on:2015-06-25Degree:MasterType:Thesis
Country:ChinaCandidate:Y N E A B D A N A y n u r A b Full Text:PDF
GTID:2180330431991828Subject:Zoology
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In recent years, illegal hunting, killing, abusing, smuggling precious animals,illegal acquisition, transportation, sale endangered wild animals and animal productbehaviors have been frequently occuring in our life, which makes the animal survivalthreatened. Therefore, it is we animal molecular biology researchers agog task to finda method to identify unknown species and to put forward effective protection strategy.DNA Barcoding have been becoming a hot spot that widely used in biologicaltaxonomy development in recent years. By this technology, compared with the knownspecies sequence recorded in Genebank, the Mitochondrial DNA (mtDNA) Ⅰcytochrome C oxidase subunit (COI) sequence of unknown species been identifed andand also able to discover a new species, rapid analysis and recognition.DNA barcoding technics have been widely used in conduct studies on fish,insects, lepidoptera, but studies about mammalian COI gene is not so common inxinjiang. For the sake of fill the lack of field using COI gene for mammalian speciesidentification, to testify the species identification international universal primers COIgene can identify xinjiang mammals or not, and also to establish a rapid, accurate,reliable mammal species identification platform and to fill the GeneBank of xinjianguyghur autonomous region animals cytochrome oxidase gene, the research identifyseven families,16species of xinjiang mammals using combination of COI genesequencing and restriction enzyme method. The result shows that:1.Using the international universal primers COI gene can be amplified inxinjiang mammals and been analysed in this research, amplification length is654bp~710bp these length completely meet requirements of the DNA barcode technology.Average A+T contents of all sequences are higher than the average G+C content,A+T content of roe deer was the highest as61.6%%, G+C content lowest as38.4%,snow leopard, A+T content of53.3%was the lowest, G+C content46.7%was thehighest, there is a clear base bias, this result according with the characteristics ofmitochondrial DNA base composition. 2.All of The62samples were analysed in this study and results showed36haplotype, and from36haplotypes found275variable sites, accounting for43.9%ofthe total sequence, including62singleton polymorphic site,213Parimonyinformation sites, in which not found singleton polymorphic site four variants, found35Parimony information sites four variants, this result explain the rich polymorphismin xinjiang mammals.3.The phylogenetic tree constructed by Neighbor joining and MaximumEvolution method get the same topology structure, each branchs contribution is veryclear, each haplotypes spreaded at single branchs,result is TRUE.4.Using the restriction enzymes Tru Ⅱcan cut COI gene sequences differentfragments of the16mammal species analysed in this study, restriction fragments wereOvis ammon491bp, Gazella subgutturosa629bp, Ovis aries470bp, Pantholopshodgsoni534bp, Pseudois nayaur457bp, Capra sibirica554bp, Procaparapicticaudata600bp,Bos grunniens491bp, Bos primigenius taurus599bp, Equuskiang414bp, Lepus timidus597bp, Uncia uncia433bp, Cervus elaphussongaricus,476bp, Capreolus capreolus396bp, Sus scrofa468bp, dog497bp, the cutpiece of the differences between species and is not obvious, such as Procaparapicticaudata and Bos primigenius Taurus, Ovis ammon and Bos grunniens have thesame enzyme loci. From the results, we learn that by the DNA barcoding technologywe can identify species accurately, but the Restriction enzyme digestion method canbe used as an auxiliary parameter for species identification. just by digestion resultsare not reliable, easy to be blending.
Keywords/Search Tags:Species identification, mtDNA, Cytochrome oxidase gene (CO1), Wild mammals, Xinjiang
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