臺灣博碩士論文加值系統

肌肉內脂肪(intramuscular fat, IMF)含量是決定豬肉品質主要因素之一，且具有高度遺傳的特性。脂肪酸結合蛋白(fatty acid-binding protein, FABP)是細胞內脂肪酸的輸送者，負責運送脂肪酸到貯存或能量產生的位置。由於此生理特性，FABP基因座成為選拔IMF含量候選基因之一。本試驗旨在應用限制片段長度多態性(restriction fragment length polymorphism, RFLP)技術，分析豬心臟型脂肪酸結合蛋白(heart fatty acid-binding protein, H-FABP)和脂肪細胞型脂肪酸結合蛋白(adipocyte fatty acid binding protein, A-FABP)基因多態性與IMF含量之關係。應用雜交肉豬311頭為試驗材料，進行IMF含量與脂肪酸組成份分析。源自血樣之DNA，以PCR進行基因片段擴增並利用RFLP技術進行基因型判定。結果顯示，肉豬H-FABP基因型之aa、DD與HH具有較高之IMF含量；而A-FABP基因型間之IMF含量無顯著差異。IMF含量分群試驗中，豬肉的飽和脂肪酸(saturated fatty acid, SFA)隨著IMF含量提高而顯著增加，但多不飽和脂肪酸(polyunsaturated fatty acid, PUFA)則隨著IMF含量提高因而顯著減少；然不同H-FABP與A-FABP基因型間之脂肪酸組成顯著不差異。比較H-FABP基因多態性對血清中脂質相關之血液生化值變化影響，結果顯示不同HaeIII與MspI基因型豬隻之血清中三酸甘油酯、膽固醇、HDL及LDL含量皆無顯著之差異。但比不同HinfI基因型豬隻時，發現HH基因型豬隻較hh型者具顯著較低之三酸甘油酯含量。綜合比較H-FABP基因型與肉中飽和脂肪酸含量發現，具高IMF含量之基因型豬隻其肉中飽和脂肪酸並未顯著地較低IMF含量者為高，故預測未來進行高IMF含量豬隻選拔時，將不致影響消費者之健康。

Intramuscular fat (IMF) content, which is highly heritable, is one of the major determinants related to pork eating quality. Fatty acid-binding proteins (FABP) play as intracellular transporters of fatty acids, which deliver fatty acids to either fat deposit sites or energy production cycle. Due to the physiological properties, fatty acid-binding protein loci are recognized as the candidate genes of IMF content related traits. Within this study, the restriction fragment length polymorphism (RFLP) techniques were used to genotype both polymorphisms of heart and adipocyte fatty acid-binding protein (H-FABP and A-FABP) genes and the associations with intramuscular fat content in pig. Data obtained from 311 hybrid hogs were used for IMF content and fatty acid compositions analysis. The DNA from blood samples was amplified by PCR and genotyped using RFLP techniques. Results indicated that hybrid hogs with aa, DD and HH H-FABP genotype had higher IMF content than those with other genotypes. In addition, no significant differences of IMF content among genotypes of A-FABP were observed in the study. The association between H-FABP genotypes and fatty acid composition was also evaluated. Results showed the higher IMF content pork was significantly associated with the higher level of saturated fatty acid (SFA) and the lower level of polyunsaturated fatty acid (PUFA). Furthermore, no significant difference of fatty acid compositions was found when H-FABP and A-FABP genotypes were compared. Comparisons of clinical blood biochemistry components related with lipids among hog H-FABP genotypes indicated that no significant difference was found in triglycerides, cholesterol, HDL or LDL levels when HaeIII and MspI analysis were evaluated, but pigs with HH genotype of HinfI analysis had lower triglycerides level than that of hh gentoype one. It implied that selecting pigs with high IMF content was not associated with high level of SFA when both H-FABP genotypes and SFA contents were considered. Therefore, it is feasible to develop a selection program for high IMF content pork production with no deterioration in consumers’ health in pig industry.

中文摘要 I
英文摘要 III
誌謝 V
目錄 VI
圖索引 IX
表索引 X
壹、前言 1
貳、文獻探討 2
一、與豬隻肥胖度性狀相關之基因 2
二、脂肪酸結合蛋白基因剔除之研究 6
三、脂肪酸結合蛋白基因之多態性 9
四、豬脂肪細胞型脂肪酸結合蛋白基因之相關研究 11
五、肉中脂肪酸組成對肉質之影響 13
參、材料與方法 19
一、樣品來源 19
二、血液樣品抽取 19
三、血液樣品基因組DNA抽取 20
四、DNA定量 23
五、引子之設計 23
六、聚合酶連鎖反應擴增目標片段 24
七、限制片段長度多態性分析 26
八、豬H-FABP與A-FABP基因PCR產物純化 28
九、豬H-FABP與A-FABP基因片段PCR產物之核苷酸定序分析及序列比對 28
十、豬隻里肌肉粗脂肪萃取 29
十一、脂肪酸組成之分析 31
十二、血清中三酸甘油酯、膽固醇、高密度脂蛋白膽固醇與低密度脂蛋白膽固醇之測定 33
十三、統計分析 37
肆、結果與討論 38
一、豬心臟型脂肪酸結合蛋白基因PCR-RFLP之多態性分析 38
二、豬脂肪細胞型脂肪酸結合基因PCR-RFLP多態性分析 39
三、豬緊迫敏感基因之多態性分析 45
四、H-FABP與A-FABP基因多態性對於肌肉內脂肪含量之影響 47
五、不同肌內脂肪含量對於脂肪酸組成之影響 49
六、H-FABP基因多態性與脂肪酸組成之關係 50
七、A-FABP基因多態性與脂肪酸組成之關係 56
八、H-FABP基因多態性對於血清中脂質含量之影響 56
九、選拔H-FABP基因型對於子代肌內脂肪含量之影響 59
伍、結論 61
參考文獻 62
作者簡介 67

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