本文選題：雞　切入點(diǎn)：akirin2基因　出處：《哈爾濱師范大學(xué)》2015年碩士論文

【摘要】：肌肉發(fā)育和抗病性狀相關(guān)基因的篩選和鑒定,對(duì)于雞的品種改良和育種具有重要的理論參考和利用價(jià)值。NF-κB是調(diào)控免疫應(yīng)答和肌肉分化過程的重要轉(zhuǎn)錄因子,AKIRIN2通過間接作用NF-κB復(fù)合物,正向調(diào)控包括IL-6在內(nèi)的多種基因的轉(zhuǎn)錄,并且akirin2基因與牛的肌內(nèi)脂肪含量和禽類肌纖維的發(fā)育存在相關(guān)性;而miR-155是一個(gè)NF-κB下游調(diào)控的非編碼小RNA,不但參與調(diào)控髓系細(xì)胞的增殖、活化和T、B細(xì)胞的分化,而且miR-155可以通過靶向肌肉發(fā)育相關(guān)基因而調(diào)控成肌細(xì)胞的分化。本研究選擇與肌肉發(fā)育和免疫相關(guān)的靶基因akirin2和miR-155,分析其在不同品系雞中的序列信息和表達(dá)分布特性,以期為雞的品系改良和分子育種相關(guān)候選基因的篩選提供理論參考。主要研究方法和實(shí)驗(yàn)結(jié)果如下:新城疫弱毒苗免疫雛雞的實(shí)驗(yàn)結(jié)果表明:免疫后7天時(shí),akirin2在肝中高表達(dá),其余組織表達(dá)變化不明顯;免疫后14天時(shí),akirin2在法氏囊中高表達(dá),在肝中表達(dá)下調(diào)。研究結(jié)果暗示akirin2不僅參與了天然免疫反應(yīng),而且對(duì)雛雞的適應(yīng)性免疫也具有重要作用。不同品系雞akirin2序列比較與表達(dá)特性分析結(jié)果表明:在四個(gè)品系雞akirin2編碼區(qū)的5'端存在4個(gè)SNP位點(diǎn),這4個(gè)堿基突變導(dǎo)致相應(yīng)位點(diǎn)三個(gè)氨基酸殘基的改變。結(jié)構(gòu)預(yù)測(cè)分析并未發(fā)現(xiàn)AKIRIN2蛋白二級(jí)結(jié)構(gòu)的差異,但有幾處蛋白結(jié)合位點(diǎn)的位置和數(shù)量發(fā)生了細(xì)微的改變,推測(cè)對(duì)AKIRIN2蛋白的表達(dá)和活性可能產(chǎn)生一定的影響。同一品系不同日齡雞的表達(dá)分析結(jié)果顯示,akirin2在各組織中廣泛表達(dá),在法氏囊、脾、血液等免疫組織中表達(dá)均為較高水平,隨著日齡的增長(zhǎng),akirin2在肌肉中的表達(dá)呈現(xiàn)出了降低的趨勢(shì)。不同品系同一日齡雞的表達(dá)分析結(jié)果顯示,akirin2在肌肉、心臟、肌胃和胸腺中的表達(dá)均較低,在血液、大腸、腺胃和肝臟中的表達(dá)均較高,但是在兩個(gè)品系雞的脂肪和脾等組織表現(xiàn)出了表達(dá)水平上的差異。miR-155的序列分析以及組織表達(dá)譜結(jié)果表明:三個(gè)品系雞pri-miR-155的序列長(zhǎng)度一致、序列無(wú)堿基缺失和突變。成熟miR-155在脊椎動(dòng)物各綱中高度保守,同源性高達(dá)100%,而pre-mi R-155的同源性低于成熟miR-155。miR-155在三個(gè)品系雞的各組織中廣泛表達(dá),無(wú)組織特異性,但在免疫相關(guān)組織中表達(dá)較高,而在肌肉組織中表達(dá)較低。
[Abstract]:Screening and identification of genes related to muscle development and disease resistance, NF- 魏 B is an important transcription factor for regulating immune response and muscle differentiation. AKIRIN2 regulates transcription of many genes, including IL-6, through indirect action of NF- 魏 B complex. Akirin2 gene is associated with the content of intramuscular fat and the development of poultry muscle fiber, and miR-155 is a non-coding small RNAs regulated downstream of NF- 魏 B, which not only participates in the regulation of proliferation, activation and differentiation of myeloid cells. Moreover, miR-155 can regulate the differentiation of myoblasts by targeting the genes associated with muscle development. In this study, the target genes akirin2 and miR-155, which are related to muscle development and immunity, were selected to analyze the sequence information and expression distribution of these genes in different strains of chicken. The main research methods and experimental results are as follows: the experimental results of Newcastle disease attenuated vaccine immunized chicks showed that: 7 days after immunization, the expression of Aakirin2 in the liver was high. There was no significant change in the expression of akirin2 in the other tissues, and the expression of kirin2 was highly expressed in bursa of Fabricius and down-regulated in liver 14 days after immunization. The results suggested that akirin2 was not only involved in innate immune response, but also in the liver. The results of akirin2 sequence comparison and expression analysis showed that there were four SNP loci at the 5 'end of akirin2 coding region in four strains of chickens. These four base mutations led to the change of three amino acid residues at the corresponding site. No difference in secondary structure of AKIRIN2 protein was found in the structural prediction analysis, but there were slight changes in the position and number of several protein binding sites. The expression and activity of AKIRIN2 protein may be affected by the same strain of different day-old chickens. The results showed that the expression of AKIRIN2 2 was widely expressed in all tissues, and the expression level was higher in immune tissues such as bursa of Fabricius, spleen, blood and so on. With the age of day, the expression of anakirin2 in muscle showed a decreasing trend. The results of expression analysis of different strains of chicken of the same day showed that the expression of pakirin2 was lower in muscle, heart, muscle stomach and thymus, and was found in blood and large intestine. The expression of adenogastric and liver was higher, but the sequence analysis and the tissue expression profile of the difference in the expression level of fat and spleen showed that the sequence length of pri-miR-155 was the same among the three strains of chicken. The homology of pre-mi R-155 was lower than that of mature miR-155.miR-155 in all tissues of three strains of chicken. However, the expression was higher in immune-related tissues and lower in muscle tissues.
【學(xué)位授予單位】：哈爾濱師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S831

【相似文獻(xiàn)】

相關(guān)期刊論文 前1條

1 曹金鎖;柳超;蔣朋飛;趙振華;陳新雨;張德禮;;豬akirin2基因的克隆與真核表達(dá)[J];中國(guó)獸醫(yī)雜志;2010年08期

相關(guān)碩士學(xué)位論文 前1條

1 常楊;不同品系雞akirin2和miR-155的克隆與表達(dá)特性分析[D];哈爾濱師范大學(xué);2015年

，

本文編號(hào)：1691847