猪血脂性状与胴体品质候选基因表达、多态检测及其相关性分析

发布时间：2018-05-01 19:57

本文选题：猪 + 血脂性状　；参考：《安徽农业大学》2016年硕士论文

【摘要】：本研究选用美系大白猪和安徽地方猪种圩猪作为瘦肉型和脂肪型猪对照对比模型,研究对比猪种间血脂性状、胴体和肉质性状的差异;选取血液胆固醇代谢途径中重要的功能基因:载脂蛋白(APOE)基因、胆固醇酯转运蛋白(CETP)基因和3-羟基-3-甲基戊二酸单酰辅酶A还原酶(HMGCR)基因为候选基因,采用PCR-SSCP技术检测APOE、CETP和HMGCR基因部分区段的遗传变异,运用SPSS软件最小二乘分析法检测该遗传变异与血脂性状的相关性,检测突变位点对候选基因在肌肉和肝脏组织表达的遗传效应;并运用SPSS软件GLM程序比较分析肝脏和背最长肌APOE、CETP和HMGCR mRNA的表达水平与猪胴体、肉质性状的相关性。以期筛选出重要的功能性多态位点作为新的遗传分子标记,以探讨利用该候选基因作为分子标记运用于猪遗传育种的可行性,为安徽地方猪种血脂及肉质性状的遗传改良提供科学根据。实验结果如下:(1)圩猪和大白猪的血脂、胴体、肉质性状的比较中,TC、TG、HDL-C、LDL-C、背膘厚、背最长肌肌肉胆固醇含量、IMF、肌内脂肪酸中硬脂酸含量均极显著高于大白猪(P0.01),十六碳烯酸含量显著高于大白猪(P0.05),亚油酸含量是极显著低于大白猪(P0.01)。圩猪血脂、胴体、肉质性状的相关性结果显示,各血脂性状之间显著正相关,血脂性状与胴体、肉质性状之间均存在不同程度的显著正相关,但与肌肉脂肪酸中亚油酸含量呈显著负相关。大白猪血脂、胴体、肉质性状的相关性结果显示,各血脂性状之间均存在显著正相关,血脂性状与胴体、肉质性状之间存在不同程度的显著正相关,与肌肉脂肪酸中亚油酸含量呈显著负相关。(2)圩猪APOE、CETP和HMGCR基因在心脏等不同组织中均有不同程度的表达,其中肝脏表达量最高,且背最长肌和背脂中均有较高程度的表达。(3)圩猪、安庆六白和大白猪APOE基因上没有检测到突变;圩猪、安庆六白和大白猪CETP基因第1内含子853碱基处发生G→T的突变位点(g.853 GT),在858碱基处存在T→C的突变位点(g.858 TC);圩猪、安庆六白和大白猪HMGCR基因在第1内含子6380碱基处存在A→C的突变位点(g.6380 AC);在第4内含子8409碱基处存在A→G的突变位点(g.8409 AG);在第11外显子14295碱基处存在T→C的错义突变位点(g.14295 TC),该核苷酸位点的突变引起氨基酸从甲硫氨酸转变为苏氨酸;在第18内含子20060碱基处存在C→T的突变位点(g.20060 CT);在第19外显子20529碱基处存在G→A的同义突变位点(g.20529 GA)。通过酶切还发现在第9外显子上存在A→G的同义突变位点(c.807AG)。圩猪和大白猪中CETP基因两个突变位点的不同基因型个体中,TC、HDL-C和LDL-C含量差异显著,TG含量差异不显著;圩猪和大白猪中HMGCR基因上3个外显子和1个内含子突变位点的不同基因型个体中,TC、HDL-C和LDL-C含量差异显著,TG含量差异不显著。且HMGCR基因mRNA在c.807AG不同基因型个体上的肝脏和肌肉表达也差异显著。(4)圩猪肝脏APOE、CETP mRNA表达量与背膘厚显著负相关,CETP mRNA表达量与背最长肌肌肉脂肪酸中的硬脂酸含量显著正相关,HMGCR基因表达与背膘厚显著正相关,与肌肉脂肪酸中的硬脂酸含量呈显著负相关;圩猪背最长肌APOE、CETP、HMGCR mRNA表达量与背膘厚、肌肉胆固醇、肌内脂肪、肌内脂肪酸中的十六碳烯酸和油酸含量呈显著正相关,与肌肉脂肪酸中的硬脂酸和亚油酸含量呈显著负相关。以上结果提示:猪血脂代谢与体内脂肪沉积密切相关,猪血脂性状的高低影响到肌内脂肪含量及其脂肪酸、胆固醇组成,对猪肉品质的形成至关重要。CETP基因g.853GT和g.858 TC可以作为猪血脂性状候选基因遗传标记;HMGCR基因g.14295 TC、g.20060 CT、g.20529 GA和c.807AG可以作为猪血脂性状候选基因遗传标记。猪血脂代谢候选功能基因可能在血脂性状和胴体、肉质性状相互影响的形成过程中发挥重要作用。
[Abstract]:In this study, a comparative model of lean and Anhui local pigs was used as a lean and fatty pig control model to compare the characteristics of blood lipids, carcass and meat quality in pigs, and to select the important functional genes in the blood cholesterol metabolism pathway: apolipoprotein (APOE) gene, cholesteryl ester transporter (CETP) gene and 3- hydroxyl The gene -3- methylglutaric acid monoyl coenzyme A reductase (HMGCR) gene is a candidate gene. The genetic variation of partial segments of APOE, CETP and HMGCR genes was detected by PCR-SSCP technique. The correlation between the genetic variation and blood lipid traits was detected by the least square analysis of SPSS software, and the expression of the mutant loci was detected in the expression of the candidate genes in the muscle and liver tissues. The SPSS software GLM program was used to compare the correlation between the expression level of APOE, CETP and HMGCR mRNA in the liver and the longest muscle of the dorsi, and the correlation between the porcine carcass and the meat quality traits. The important functional polymorphic loci were selected as a new genetic marker to explore the use of the candidate gene as a molecular marker in pig genetic breeding. The experimental results are as follows: (1) the blood lipid, carcass and meat quality of polder and white pigs were compared, TC, TG, HDL-C, LDL-C, back fat thick, the muscle cholesterol content of the longest muscle, IMF, and the content of stearic acid in intramuscular fatty acids were significantly higher than that of white. The content of sixteen cartonic acid was significantly higher than that of the white pig (P0.05), and the content of linoleic acid was significantly lower than that of the big white pig (P0.01). The correlation of blood lipids, carcass and meat quality in polder pigs showed that there was significant positive correlation between the lipid traits and the significant positive correlation between the lipid traits and the carcass and meat quality traits, but with the muscles. The correlation between fatty acid and fatty acids showed significant negative correlation. The correlation between fat, carcass and meat quality showed that there was significant positive correlation between the lipid traits and the significant positive correlation between the lipid traits and carcass, meat quality traits, and the significant negative correlation with the fatty acid content of the fatty acid. (2) the polder pigs APOE, C ETP and HMGCR genes were expressed in different degrees in different tissues of the heart, with the highest expression in the liver, and high expression in the longest and back fat. (3) the polder pigs, the Anqing six white and the big white pig APOE gene did not detect the mutation; the polder pig, the Anqing six white and the big white pig CETP gene first intron 853 bases occurred G The mutation site of T (g.853 GT) has a mutation site of T to C (g.858 TC) at the 858 base. The polder pig, the HMGCR gene of Anqing six white and the big white pig has the mutation site of A to C in the 6380 base of the intron first (g.6380 AC), and there is a mutation site at the 8409 base of the fourth intron, and there is a 14295 base at the eleventh exon 14295 base. C's missense mutation site (g.14295 TC), the mutation of the nucleotide site causes the transformation of amino acids from methionine to threonine, and there is a mutation site (g.20060 CT) at the 20060 base of the intron eighteenth (g.20060 CT), and there is a synonymous mutation point of G to A (g.20529 GA) at the 20529 base of exon nineteenth (g.20529 GA). It is also found on the ninth exons by the enzyme cut. There is a synonymous mutation site (c.807AG) of A to G. Among the different genotype individuals of the two mutation sites of the CETP gene in polder pigs and big white pigs, there are significant differences in the content of TC, HDL-C and LDL-C, and there is no significant difference in TG content; of the 3 exons and 1 intron mutant loci in the polder and white pigs, TC, HDL-C, and LDL-C. There was no significant difference in content, and there was no significant difference in TG content. And the expression of HMGCR gene mRNA in the liver and muscles of different genotypes of c.807AG was also significant. (4) the expression of APOE in the liver of the polder pigs, CETP mRNA expression was significantly negatively correlated with the backfat thickness, and the mRNA expression of CETP was significantly correlated with the stearic acid content in the muscle fatty acid of the dorsal longest muscle, HMGCR gene. The expression was significantly positively correlated with backfat thickness, and was negatively correlated with stearic acid content in muscle fatty acids. The expression of APOE, CETP, HMGCR mRNA in the longest muscle of polder piglets was significantly correlated with the thickness of back fat, muscle cholesterol, intramuscular fat, sixteen ARIC acid and oleic acid content in intramuscular fatty acids, with stearic acid and linoleic acid in muscle fatty acids. The above results suggest that the blood lipid metabolism in pigs is closely related to the body fat deposition. The high and low lipid content of pigs affects the content of intramuscular fat and the composition of fatty acids and cholesterol. The formation of.CETP gene g.853GT and g.858 TC can be used as a genetic marker for the candidate gene of porcine blood lipid; HMGCR base G.14295 TC, g.20060 CT, g.20529 GA and c.807AG can be used as genetic markers for the candidate gene for porcine blood lipid traits. The candidate gene for pig lipid metabolism may play an important role in the formation of blood lipid traits and the interaction of carcass and meat quality traits.

【学位授予单位】：安徽农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S828

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