本文選題：雉科鳥類 + BF　； 參考：《東北林業(yè)大學》2016年碩士論文

【摘要】：目前鳥類各種傳染性疾病對種群數(shù)量構(gòu)成嚴重威脅,因此鳥類抗病性研究已成為熱點。BF基因和Mx基因均是自然選擇作用下產(chǎn)生的抗病性基因,并且這兩種基因的多態(tài)性在鳥類抵抗感染和自身免疫疾病方面起著至關重要的作用,同時也是禽類抗病育種的有效候選基因。本研究以黃腹角雉等10種雉科鳥類為研究對象,通過直接測序或平端克隆的方法獲得黃腹角雉3個個體的BF基因外顯子2-8序列和5種雉科鳥類的BF基因外顯子2、3序列以及10種雉科鳥類Mx基因外顯子13和外顯子14序列。并探討這兩種基因的多態(tài)性及進化,為進一步研究鳥類BF和Mx基因的抗病性奠定理論基礎,結(jié)果如下：1.獲得黃腹角雉3個個體的BF基因部分序列,包括7個外顯子(外顯子2-8)和6個內(nèi)含子(內(nèi)含子2-7),確定了9個單倍型,序列長1803～1828bp,編碼區(qū)長1010bp,共編碼335個氨基酸。9個單倍型中,核苷酸變異位點最豐富的區(qū)域位于al(exon2編碼)和a2 (exon3編碼)結(jié)構(gòu)域構(gòu)成的抗原結(jié)合區(qū)。2.從雉科5種鳥類的25個個體的125條有效的BF exon2、3完整編碼區(qū)克隆序列中確定了20個單倍型,序列長544bp,共編碼181個氨基酸,核苷酸變異位點192個,其中簡約信息位點126個。3.獲得雉科10種鳥類的Mx基因外顯子13和外顯子14序列,其中外顯子13的40條序列中確定了18個單倍型,外顯子14的58條序列中確定了13個單倍型,結(jié)果顯示外顯子14更加保守。4.BF基因的α1結(jié)構(gòu)域抗原結(jié)合區(qū)(PBR)受正選擇作用,而非抗原結(jié)合區(qū)(non-PBR)受凈化選擇作用；α2結(jié)構(gòu)域,無論是抗原結(jié)合區(qū)(PBR)還是非抗原結(jié)合區(qū)(non-PBR)皆受到凈化選擇作用,但抗原結(jié)合區(qū)的非同義替換率均大于非抗原結(jié)合區(qū)。Mx基因的GED區(qū)以及位于GED區(qū)控制Mx蛋白抗病毒活性的S631N突變位點則均受凈化選擇作用。5.BF基因和Mx基因中皆存在屬內(nèi)與屬間共享單倍型,并且BF基因所構(gòu)建的進化樹顯示出此基因的跨物種進化現(xiàn)象。6.本實驗獲得的雉科10種鳥類Mx基因第631位氨基酸均為絲氨酸(Ser),所以推測本研究的這十種野生雉科鳥類群體皆屬于易感群體。
[Abstract]:At present, a variety of infectious diseases in birds pose a serious threat to the population, so the study of bird disease resistance has become a hot spot. BF gene and Mx gene are the resistance genes produced by natural selection.The polymorphism of these two genes plays an important role in bird resistance to infection and autoimmune diseases, and is also an effective candidate gene for avian disease resistance breeding.In this study, 10 species of pheasants, such as the Yellow Pheasant, were selected as the research objects.The exon 2-8 sequence of BF gene in three individuals of the pheasant and the sequence of exon 2C3 of BF gene in 5 species of pheasants, and the sequence of exon 13 and exon 14 of Mx gene in 10 species of pheasants were obtained by direct sequencing or flat-end cloning.The polymorphism and evolution of the two genes were discussed, which laid a theoretical foundation for further study on the resistance of BF and Mx genes in birds. The results are as follows: 1.The BF gene partial sequences of three individuals of the pheasant were obtained, including 7 exons (exon 2-8) and 6 introns (intron 2-7). Nine haplotypes were identified, the length of the sequence was 1803 ~ 1828 BP, and the coding region was 1010 BP, encoding a total of 335 amino acids.The most abundant regions of nucleotide variation are located in the antigen-binding region of al(exon2 coding) and a2 + exon3) domain.Twenty haplotypes, encoding 181 amino acids and 192 nucleotide mutation sites, were identified from 125 effective BF exon 2N 3 complete coding regions from 25 individuals of 5 species of the pheasant family, with a total length of 544bp. among them, the reduced information sites were 126. 3.Exon 13 and exon 14 sequences of Mx gene were obtained from 10 species of pheasant family. Among them, 18 haplotypes were identified in 40 exon 13 sequences and 13 haplotypes were identified in 58 exon 14 sequences.The results showed that exon 14 was more conserved. 4. The 偽 1 domain antigen binding region (PBR) of BF gene was positively selected, but the non-antigen binding region (non-PBR) was purified and the 偽 2 domain was purified.Both antigen-binding region (PBR) and non-antigen binding region (non-PBR) were purified and selected.However, the non-synonymous substitution rate of antigen binding region was higher than that of GED region of non-antigen binding region. Mx gene and S631N mutation site located in GED region which controlled the antiviral activity of Mx protein. 5. BF gene and Mx gene were all purified and selected.Shared haplotypes within and between genera,Furthermore, the evolutionary tree constructed by BF gene showed the cross species evolution phenomenon of BF gene.The amino acids at position 631 of Mx gene of 10 species of pheasants in this study were all serine serine, so we speculate that all the ten species of wild birds in this study belong to susceptible populations.
【學位授予單位】：東北林業(yè)大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：Q953

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本文編號：1763622