本文選題：豬 + 血脂性狀�。� 參考：《安徽農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：本研究選用美系大白豬和安徽地方豬種圩豬作為瘦肉型和脂肪型豬對(duì)照對(duì)比模型,研究對(duì)比豬種間血脂性狀、胴體和肉質(zhì)性狀的差異;選取血液膽固醇代謝途徑中重要的功能基因:載脂蛋白(APOE)基因、膽固醇酯轉(zhuǎn)運(yùn)蛋白(CETP)基因和3-羥基-3-甲基戊二酸單酰輔酶A還原酶(HMGCR)基因?yàn)楹蜻x基因,采用PCR-SSCP技術(shù)檢測(cè)APOE、CETP和HMGCR基因部分區(qū)段的遺傳變異,運(yùn)用SPSS軟件最小二乘分析法檢測(cè)該遺傳變異與血脂性狀的相關(guān)性,檢測(cè)突變位點(diǎn)對(duì)候選基因在肌肉和肝臟組織表達(dá)的遺傳效應(yīng);并運(yùn)用SPSS軟件GLM程序比較分析肝臟和背最長(zhǎng)肌APOE、CETP和HMGCR mRNA的表達(dá)水平與豬胴體、肉質(zhì)性狀的相關(guān)性。以期篩選出重要的功能性多態(tài)位點(diǎn)作為新的遺傳分子標(biāo)記,以探討利用該候選基因作為分子標(biāo)記運(yùn)用于豬遺傳育種的可行性,為安徽地方豬種血脂及肉質(zhì)性狀的遺傳改良提供科學(xué)根據(jù)。實(shí)驗(yàn)結(jié)果如下:(1)圩豬和大白豬的血脂、胴體、肉質(zhì)性狀的比較中,TC、TG、HDL-C、LDL-C、背膘厚、背最長(zhǎng)肌肌肉膽固醇含量、IMF、肌內(nèi)脂肪酸中硬脂酸含量均極顯著高于大白豬(P0.01),十六碳烯酸含量顯著高于大白豬(P0.05),亞油酸含量是極顯著低于大白豬(P0.01)。圩豬血脂、胴體、肉質(zhì)性狀的相關(guān)性結(jié)果顯示,各血脂性狀之間顯著正相關(guān),血脂性狀與胴體、肉質(zhì)性狀之間均存在不同程度的顯著正相關(guān),但與肌肉脂肪酸中亞油酸含量呈顯著負(fù)相關(guān)。大白豬血脂、胴體、肉質(zhì)性狀的相關(guān)性結(jié)果顯示,各血脂性狀之間均存在顯著正相關(guān),血脂性狀與胴體、肉質(zhì)性狀之間存在不同程度的顯著正相關(guān),與肌肉脂肪酸中亞油酸含量呈顯著負(fù)相關(guān)。(2)圩豬APOE、CETP和HMGCR基因在心臟等不同組織中均有不同程度的表達(dá),其中肝臟表達(dá)量最高,且背最長(zhǎng)肌和背脂中均有較高程度的表達(dá)。(3)圩豬、安慶六白和大白豬APOE基因上沒有檢測(cè)到突變;圩豬、安慶六白和大白豬CETP基因第1內(nèi)含子853堿基處發(fā)生G→T的突變位點(diǎn)(g.853 GT),在858堿基處存在T→C的突變位點(diǎn)(g.858 TC);圩豬、安慶六白和大白豬HMGCR基因在第1內(nèi)含子6380堿基處存在A→C的突變位點(diǎn)(g.6380 AC);在第4內(nèi)含子8409堿基處存在A→G的突變位點(diǎn)(g.8409 AG);在第11外顯子14295堿基處存在T→C的錯(cuò)義突變位點(diǎn)(g.14295 TC),該核苷酸位點(diǎn)的突變引起氨基酸從甲硫氨酸轉(zhuǎn)變?yōu)樘K氨酸;在第18內(nèi)含子20060堿基處存在C→T的突變位點(diǎn)(g.20060 CT);在第19外顯子20529堿基處存在G→A的同義突變位點(diǎn)(g.20529 GA)。通過酶切還發(fā)現(xiàn)在第9外顯子上存在A→G的同義突變位點(diǎn)(c.807AG)。圩豬和大白豬中CETP基因兩個(gè)突變位點(diǎn)的不同基因型個(gè)體中,TC、HDL-C和LDL-C含量差異顯著,TG含量差異不顯著;圩豬和大白豬中HMGCR基因上3個(gè)外顯子和1個(gè)內(nèi)含子突變位點(diǎn)的不同基因型個(gè)體中,TC、HDL-C和LDL-C含量差異顯著,TG含量差異不顯著。且HMGCR基因mRNA在c.807AG不同基因型個(gè)體上的肝臟和肌肉表達(dá)也差異顯著。(4)圩豬肝臟APOE、CETP mRNA表達(dá)量與背膘厚顯著負(fù)相關(guān),CETP mRNA表達(dá)量與背最長(zhǎng)肌肌肉脂肪酸中的硬脂酸含量顯著正相關(guān),HMGCR基因表達(dá)與背膘厚顯著正相關(guān),與肌肉脂肪酸中的硬脂酸含量呈顯著負(fù)相關(guān);圩豬背最長(zhǎng)肌APOE、CETP、HMGCR mRNA表達(dá)量與背膘厚、肌肉膽固醇、肌內(nèi)脂肪、肌內(nèi)脂肪酸中的十六碳烯酸和油酸含量呈顯著正相關(guān),與肌肉脂肪酸中的硬脂酸和亞油酸含量呈顯著負(fù)相關(guān)。以上結(jié)果提示:豬血脂代謝與體內(nèi)脂肪沉積密切相關(guān),豬血脂性狀的高低影響到肌內(nèi)脂肪含量及其脂肪酸、膽固醇組成,對(duì)豬肉品質(zhì)的形成至關(guān)重要。CETP基因g.853GT和g.858 TC可以作為豬血脂性狀候選基因遺傳標(biāo)記;HMGCR基因g.14295 TC、g.20060 CT、g.20529 GA和c.807AG可以作為豬血脂性狀候選基因遺傳標(biāo)記。豬血脂代謝候選功能基因可能在血脂性狀和胴體、肉質(zhì)性狀相互影響的形成過程中發(fā)揮重要作用。
[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.

【學(xué)位授予單位】：安徽農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S828

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