本文關(guān)鍵詞：ACSL5基因?qū)θ馀ｋ伢w性狀及脂肪合成通路甘油三酯的影響　出處：《吉林大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： ACSL5 肉牛 胴體組成性狀 牛前體脂肪細(xì)胞 甘油三酯 脂肪代謝通路

【摘要】：長鏈脂酰輔酶A合成酶5(ACSL5)是ACSLs家族中的重要一員,是動物機體脂肪酸β氧化和甘油三酯合成、代謝過程中的關(guān)鍵酶。但ACSL5基因?qū)θ馀Ｖ舅岷铣伞⒅境练e和胴體性狀是否亦具有相同的影響作用尚不清楚。本實驗通過肉牛群體和細(xì)胞水平對ACSL5基因調(diào)控脂肪代謝作用進行了研究分析,為進一步解析ACSL5的功能及應(yīng)用于肉牛分子輔助育種提供技術(shù)依據(jù)。本實驗以肉用牛為研究對象,測定了354頭中國西門塔爾牛的胴體與肉質(zhì)性狀,采用PCR-RFLP對該群體進行了ACSL5基因多態(tài)性檢驗,分析了其SNPs與肉牛群體胴體性狀和肉質(zhì)性狀的相關(guān)性。進而,構(gòu)建了牛ACSL5過表達載體PBI-CMV3-ACSL5和sh RNA干擾載體,采用電穿孔方法轉(zhuǎn)染牛前體脂肪細(xì)胞,解析了ACSL5基因表達水平變化(過表達和沉默后)對細(xì)胞的脂肪代謝通路中甘油三酯合成的作用。結(jié)果表明,在中國西門塔爾牛的5’側(cè)翼序列中篩查到兩個SNP位點g.33185918GA和g.33186348CT,與中國西門塔爾肉牛的胴體和肉質(zhì)性狀具有顯著的相關(guān)性。g.33185918GA位點,相比于AA純合基因型個體,中國西門塔爾牛群體中攜帶G等位基因(GA基因型或GG基因型)的個體表現(xiàn)出更高的瘤網(wǎng)皺胃重(7.48±0.91 kg)、瓣胃重(3.94±0.59 kg)、睪丸重(0.69±0.13 kg)和脂肪顏色評分(2.97±0.74,p0.05);而AA純合基因型個體表現(xiàn)出更高的排酸p H值(5.91±0.78 kg,p0.05);其中,瘤網(wǎng)皺胃重和排酸p H值均表現(xiàn)為極顯著差異(p0.01)。g.33186348CT位點,與CC純合基因型中國西門塔爾牛個體相比,攜帶T等位基因(CT基因型和TT基因型)的個體表現(xiàn)出了更高的胴體脂肪覆蓋率(50.38±20.46%)、腰部肉厚(6.89±0.87 cm)和肝臟重(6.33±1.24kg),而攜帶CC純合基因型的個體卻表現(xiàn)出更高的后蹄重(3.39±1.03 kg)、后腿長(79.87±3.98 cm)、生殖器脂肪重(0.91±0.37 kg)和胴體長(139.50±7.79 cm);其中,肝臟重、后腿重、腎臟重、睪丸重和脂肪覆蓋率均有極顯著差異(p0.01)。說明,ACSL5基因調(diào)控區(qū)域的SNPs與中國西門塔爾牛部分胴體性狀和肉質(zhì)性狀具有相關(guān)性。在牛前體脂肪細(xì)胞中進行ACSL5基因的過表達和沉默后處理后檢測細(xì)胞脂肪合成通路中甘油三酯的變化。結(jié)果表明,ACSL5基因在牛前體脂肪細(xì)胞中過表達能顯著提高細(xì)胞中甘油三酯的含量(p0.01);而當(dāng)牛前體脂肪細(xì)胞中沉默ACSL5基因表達后,細(xì)胞內(nèi)甘油三酯合成量有降低趨勢,但并不顯著(p0.05)。綜上所述,ACSL5基因調(diào)控區(qū)域發(fā)生的SNP:g.33185918GA和g.33186348CT均與中國西門塔爾肉牛部分胴體和肉質(zhì)性狀存在相關(guān)性;同時ACSL5基因?qū)εＧ绑w脂肪細(xì)胞甘油三酯的合成具有一定促進作用。研究結(jié)果可為進一步解析ACSL5的功能及應(yīng)用于肉牛分子輔助育種提供技術(shù)依據(jù)。
[Abstract]:Long chain acyl coenzyme A synthetase 5 (ACSL5) is an important member of the ACSLs family, is the animal body fatty acid oxidation and triglyceride synthesis, the key enzyme in the metabolic process. But ACSL5 gene on beef fatty acid synthesis, fat deposition and carcass traits are also have the same effect is not clear. Through the experiments of beef cattle populations and cellular levels of ACSL5 gene regulating fat metabolism were analyzed for further analysis the function and application of ACSL5 in beef cattle molecular breeding to provide technical basis. In this experiment, the beef cattle as the research object, measured 354 Chinese Simmental Cattle Carcass and meat quality traits, PCR-RFLP is used test of ACSL5 gene polymorphism of the group, to analyze the correlation between the SNPs group and beef carcass traits and meat quality. Then, constructed a bovine ACSL5 expression vector PBI-CMV3-ACSL5 and sh RNA Interference vector by electroporation into bovine preadipocytes, the expression of the ACSL5 gene (overexpression and silencing) role of triglyceride fatty metabolic pathway on cell synthesis. The results showed that in China Simmental 5 flanking sequence screened two SNP loci g.33185918GA and g.33186348CT, with correlation.G.33185918GA significant sites with carcass and meat quality traits in Simmental beef cattle China, compared to the AA genotype, Chinese Simmental population carrying G allele (GA genotype or GG genotype) of the individuals showed tumor net abomasum higher weight (7.48 + 0.91 kg (3.94), heavy omasum 0.59 + kg), testis weight (0.69 + 0.13 kg) and fat color score (2.97 + 0.74, P0.05); and AA genotype showed Acid P higher H value (5.91 + 0.78 kg, P0.05); the tumor weight and net abomasum P acid H value showed significant difference (P0.01).G.33186348CT loci, and CC homozygous gene compared China Simmental individuals carrying T allele (CT and TT genotypes) individuals showed higher carcass fat coverage (50.38 + 20.46%), the waist of the flesh (6.89 + 0.87 cm) and liver weight (6.33 + 1.24kg), while carrying CC homozygous genotype showed higher after hoof weight (3.39 + 1.03 kg) and long hind legs (79.87 + 3.98 cm), genital fat weight (0.91 + 0.37 kg) and carcass length (139.50. 7.79 cm); among them, liver weight, kidney weight, leg weight, testis weight and fat coverage rate were significantly different (P0.01). The results showed that, ACSL5 gene regulatory region SNPs is associated with China Simmental some carcass traits and meat quality traits. In bovine preadipocyte cells were detected after postprocessing. Expression and gene silencing by ACSL5 Changes in fatty triglyceride synthesis pathway. The results showed that the ACSL5 gene in bovine preadipocyte overexpression can significantly improve the content of triglyceride (P0.01); and when the silencing of ACSL5 gene expression in bovine preadipocyte, intracellular triglyceride synthesis quantity has decreased, but not significant (P0.05). In summary, both the regulatory regions of ACSL5 SNP:g.33185918GA and g.33186348CT are associated with China part of Simmental Beef Cattle Carcass and meat quality traits; at the same time the synthesis of ACSL5 gene of bovine preadipocyte triglyceride has certain stimulative effect. The research results can be used for further analysis of the function and application of ACSL5 in beef cattle molecular breeding to provide technical basis.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S823

【參考文獻】

相關(guān)期刊論文 前10條

1 薩日娜;武會娟;;脂代謝通路及脂代謝異常在癌癥中的研究現(xiàn)狀[J];腫瘤防治研究;2016年10期

2 李聰;曹文廣;;CRISPR/Cas9介導(dǎo)的綿羊胎兒成纖維細(xì)胞肌肉生長抑制素基因敲除研究[J];中國畜牧獸醫(yī);2015年11期

3 滑留帥;王t，

本文編號：1418397