| Wild Argali is a superordinary wildlife resource in Xinjiang,China.The breed have many features,for example, macro-somatotype,better anti-disease,better accommodation,better toc-flesh and high lean meat rate, and so on. In order to utilize and protect wild animal resource of xinjiang, import blood lineage of Wild Argali largely, the experiment use Wild Argali ((?)),BaShenBai ((?)) or F1 ((?)) for crossing by artificial insemination fecundation The results show that they give birth to crossing sheep of F1,F2,but the anti-disease of F1,F2 is poor.While Wild Argali,BaShenBai have no disease at the same raising condition.So we presume that some disease resistance relational genes of crossing lamb appear variation.Therefore, the TLR9å'ŒISG15 disease resistance relational genes were studied by comparative genomics way,to research the mechanism of the poor anti-disease power of F1,F2 crossing lamb. In addition,It was a hot point which was in molecular biology to research the diversity of animal heredity and the evolution relation among species in the level of DNA,as the research of other animals in this feild,the research in Wild Argali which was belonged to our country had research shallowly.Therefore,we analyzed the Wild Argali mtDNA D-loop region,for the elucidation of the diversity of animal heredity and the evolution relation of Wild Argali in our country.1.Wild Argali,BaShenBai and crossing sheep ISG15 Cloning and Sequence AnalysisThe experiment aims at studying for the mRNA,conservative structural domain,3-D structure of ISG15. The ISG15 complete sequences were specificitly amplified by PCR technology.The PCR product was purified by agarose.The ISG15 complete sequences were sequenced and analysised by the method of comparative genomics. The result showed the length of ISG15 complete sequence were 2117bp,2123bp,2074bp,2121bp,respectively,in wild argali,Bashenbai,F1,F2; The Wild Argali Xinjiang ISG15 mRNA sequence had 94.7% identity with the sheep ISG15 mRNA, and cow88.8%, F1 97.1%, pig 80.6%,cat 76.6%, human 67.7%,mouse 66.5%,fish 45.5%. The Wild Argali Xinjiang ISG15 protein sequence had 99.3% identity with the sheep, and cow 89.1%, Fl 92.8%, pig 74.1%,cat 72.1%, human 63.6%,mouse 63.0%,fish 29.0%. The conserved domain of ISG15 were analysised by the method of comparative genomics. The result showed the quantity, type and the relative position of ISG15 protein motif were different in several species.ISG15 contains two domains with structural homology close to ubiquitin, and species protein 3-D structures were also conservative.Interestly, ISG15 contains one UBQ domain and one ubiquitin multifunction domain in several species, except for F1 which contains one UBQ multifunction domain. The investigation presumed that because of the difference of filial generationISG15gene in the mRNA sequence the quantity of protein conservative stuctural domains and the 3D structure between parental generartion ISG15 gene,resulted the effect of filial generation ISG15gene variancein innate immunity and anti-disease.So the anti-disease of filial generation became weak.2.Wild Argali,BaShenBai and crossing sheep TLR9 Cloning and Sequence AnalysisThe experiment aims at studying for the mRNA,conservative structural domain,3-D structure of TLR9. The TLR9 complete sequences were specificitly amplifited by PCR technology.The PCR product was purified by agarose.The TLR9 complete sequences were sequenced and analysised by the method of comparative genomics. TLR9 were 3193bp,3169bp,3103bpå'Œ3109bp.After gene predicted, The Wild Argali Xinjiang TLR9 mRNA sequence had 99.9% identity with the sheepTLR9 mRNA, and goat 99%,cow 95.2%, pig 85.1%, cat 84%, dog 83.7%, horse 83.4%, human79.4%,monkey 79.1%,mouse 73.1%. The Wild Argali Xinjiang TLR9 protein sequence had 97.7% identity with the sheep TLR9 protein, and goat97%,cow 94.6%, pig 86.2%, cat 84%, dog83.7%, horse 83.4%, human79.4%,monkey 79.1%,mouse 73.1%.The conserved domain of TLR9 were analysised by the method of comparative genomics. The result showed the quantity, type and the relative position of TLR9 protein motif were different in several species. The 3-D of TLR9 was constructived by LRR and TIL(Toll/IL IR) structural domain.The above-mentioned structural feature offered theory accordings to research TLR9 of Wild Argali (O.a.karelini),Xinjiang,China.The investigation presumed that the quantity, type,the relative position of TLR9 conservative structural domain and 3-D structure rch of TLR9 protein motif,which came from Wild Argali and BaShenBai, might occurred distinction on crossing lamb.it led to decrease shaply the anti-disease power of crossing lamb,and then it easily infected infectiousness pleuropneumonia in the immature stage of crossing lamb. 3.Wild Argali mitochondria displacement loop region Cloning and Sequence AnalysisThe experiment aims at studying Mitochondrial DNA (mtDNA) Genetic Diversity of Wild Argali.The mtDNA D-loop complete sequences of Wild Argali (O.a.karelini),Xinjiang,China were specificitly amplifited by PCR technology.The PCR product was purified by agarose.The mtDNA D-loop sequences were sequenced and analysised. The result showed the length of mtDNA D-loop complete sequence was 1070bp and 1169bp,respectively,in number one and number two.There were 7 insertion loci,50 transitions loci, lltransversion loci and 5 deletions loci 70 mutational loci which were found in two wild Argalis and transitions was the major mutational type.The mutational loci of mtDNA D-loop sequence, from number one and number two Wild Argali (O.a.karelini), were 35 and 38; mutational ratewere 3.12% and 3.25%.It showed that the Wild Argali (O.a.karelini) population has high variation in mtDNA D-loop sequence.Meanwhile,it further showed that there was plentiful genetic resource of Wild Argali Xinjiang,China. On the basis of the analysis of system evolution tree between Wild Argali and other sheep,the relation between Wild Argali and other sheep was far.In speculation Wild Argali was not the ancestor of our country domesticated sheep. |
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