本文選題：小尾寒羊　切入點(diǎn)：脂聯(lián)素基因　出處：《山東農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：脂肪對(duì)于綿羊維持正常生命活動(dòng)及生理代謝發(fā)揮重要的作用,脂肪性狀是影響綿羊肌肉品質(zhì)的一個(gè)重要的經(jīng)濟(jì)性狀。脂聯(lián)素(Adipoq)是脂肪組織分泌的一種內(nèi)源性特異蛋白質(zhì),也是一種重要的脂肪細(xì)胞因子。本試驗(yàn)克隆了小尾寒羊脂聯(lián)素基因的全長cDNA序列,并對(duì)該基因及其所編碼的蛋白、mRNA水平和蛋白水平上的表達(dá)豐度進(jìn)行了分析。主要研究結(jié)果如下:1.采用RT-PCR和RACE方法,以小尾寒羊組織為試驗(yàn)材料,克隆了小尾寒羊脂聯(lián)素基因的全長cDNA序列,獲得GenBank登錄號(hào)為KM216385。該基因cDNA全長為2616 bp,包含720 bp的開放閱讀框、76 bp的5'非編碼區(qū)(5'UTR)和1820 bp的3'非編碼區(qū)(3'UTR)。其中,開放閱讀框編碼239個(gè)氨基酸。2.生物信息學(xué)分析顯示,脂聯(lián)素基因編碼的蛋白分子量為26 kDa,等電點(diǎn)為5.88,并存在信號(hào)肽結(jié)構(gòu),能被分泌到細(xì)胞外發(fā)揮作用。在氨基酸序列中,發(fā)現(xiàn)2個(gè)蘇氨酸、3個(gè)絲氨酸和3個(gè)酪氨酸殘基存在磷酸化位點(diǎn),并發(fā)現(xiàn)存在5個(gè)O-糖基化位點(diǎn)而無N-糖基化位點(diǎn)。與其他物種的氨基酸同源性分析,證明綿羊脂聯(lián)素蛋白的氨基酸序列與山羊的相似性最高。3.檢測到Adipoq基因在小尾寒羊和杜泊羊中呈現(xiàn)出組織表達(dá)差異性。不同組織間,脂聯(lián)素基因在皮下脂肪、腎周脂肪、腹股溝脂肪、臂二頭肌、心血管組織中有表達(dá),在肝臟、腎臟、卵巢組織中未檢測到表達(dá),提示脂聯(lián)素基因的表達(dá)有一定的組織特異性。其中,該基因在三種脂肪組織中表達(dá)水平最高,極顯著高于其他組織(P0.01),但是三種脂肪組織之間的表達(dá)量無顯著差異(P0.05)。不同品種間,脂聯(lián)素基因在杜泊羊臂二頭肌組織中的表達(dá)極顯著高于小尾寒羊(P0.01)。4.檢測到Adipoq蛋白在小尾寒羊和杜泊羊中存在組織表達(dá)差異性。不同組織間,脂聯(lián)素蛋白在皮下脂肪、腎周脂肪、腹股溝脂肪、臂二頭肌、心血管組織中有表達(dá),在肝臟、腎臟組織中未檢測到表達(dá)。其中,脂聯(lián)素蛋白在三種脂肪組織中表達(dá)量最高,極顯著高于臂二頭肌和心血管(P0.01),但是三種脂肪組織之間的表達(dá)量無顯著差異(P0.05);不同品種間,脂聯(lián)素蛋白在三種脂肪組織中的表達(dá)量,小尾寒羊均高于杜泊羊,但差異均不顯著(P0.05)。杜泊羊臂二頭肌的脂聯(lián)素蛋白表達(dá)量高于小尾寒羊,而心血管中脂聯(lián)素蛋白表達(dá)水平低于小尾寒羊,但是差異也不顯著(P0.05)。綜上所述,本研究成功克隆了小尾寒羊脂聯(lián)素基因,對(duì)該基因序列及其編碼的氨基酸序列的結(jié)構(gòu)特征進(jìn)行了預(yù)測,并比較分析了小尾寒羊和杜泊羊脂聯(lián)素基因mRNA水平和蛋白水平上的組織表達(dá)譜特征,為后續(xù)研究脂聯(lián)素的功能提供了一定的依據(jù)。
[Abstract]:Fat plays an important role in maintaining normal life activity and physiological metabolism in sheep. Adiponectin (Adipoq) is an endogenous protein secreted by adipose tissue, which is an important economic trait affecting the muscle quality of sheep. In this study, we cloned the full length cDNA sequence of adiponectin gene from small tail Han sheep. The mRNA level and protein abundance of the gene were analyzed. The main results were as follows: 1. Using RT-PCR and RACE methods, the tissues of small tail Han sheep were used as experimental materials. The full-length cDNA sequence of adiponectin gene of small tail Han sheep was cloned and the GenBank accession number was KM216385.The cDNA length of the gene was 2616 BP, containing 720bp open reading frame 76 BP 5'noncoding region 5 'unencoded UTRR and 1820 BP 3'noncoding region 3' UTR. among them, Open reading frame encodes 239 amino acids. Bioinformatics analysis shows that adiponectin gene encodes protein with a molecular weight of 26 kDa and a isoelectric point of 5.88, and has a signal peptide structure that can be secreted to play an extracellular role. Two threonine, three serine and three tyrosine residues were found to have phosphorylation sites, and five O-glycosylation sites without N-glycosylation sites were found. The results showed that the amino acid sequence of sheep adiponectin had the highest similarity with goat. 3. The difference of tissue expression of Adipoq gene was found in small tail Han sheep and du Bo sheep. Adiponectin gene was found to be subcutaneous fat and perirenal fat in different tissues. The expression of adiponectin gene in inguinal fat, biceps arm muscle, cardiovascular tissue was not detected in liver, kidney and ovary tissues, suggesting that adiponectin gene expression has certain tissue specificity. The highest expression level of the gene was found in three kinds of adipose tissues, which was significantly higher than that in other tissues, but there was no significant difference in the expression levels among the three kinds of adipose tissues. The expression of adiponectin gene was significantly higher in the biceps of arm than that in the small tail Han sheep (P 0.01). 4. The difference of tissue expression of adiponectin was detected between small tail Han sheep and du Bo sheep. Adiponectin was found in subcutaneous fat and perirenal fat in different tissues. Inguinal fat, biceps arm muscle, cardiovascular tissue, no expression in liver and kidney tissue, adiponectin in three kinds of adipose tissue, adiponectin expression is the highest in the three kinds of adipose tissue, The expression of adiponectin in the three adipose tissues was significantly higher than that in the biceps arm muscle and the cardiovascular tissue, but there was no significant difference among the three adipose tissues, and the expression of adiponectin in the three kinds of adipose tissues was higher in small tail Han sheep than that in Dupo sheep. But the difference was not significant (P 0.05). The adiponectin expression in the biceps of arm was higher than that in the small tail Han sheep, but the level of adiponectin in cardiovascular was lower than that in the small tail Han sheep, but the difference was not significant (P 0.05). In this study, the adiponectin gene of small tail Han sheep was cloned successfully, and the structural characteristics of the gene and the amino acid sequence were predicted. The tissue expression profiles of adiponectin gene mRNA and protein in small tail Han sheep and Dupo sheep were compared and analyzed, which provided a basis for further study of adiponectin function.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S826

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相關(guān)期刊論文 前1條

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本文編號(hào)：1570270