本文關(guān)鍵詞：不同品種豬肌纖維組成成分及相關(guān)信號(hào)通路基因的表達(dá)研究　出處：《錦州醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 豬 品種 肉質(zhì) 肌纖維 信號(hào)通路

【摘要】：目的探索不同品種豬出生后肌纖維組成成分及相關(guān)信號(hào)通路關(guān)鍵成分的基因表達(dá)規(guī)律及種間差異,同時(shí)分析豬肉質(zhì)性狀候選基因CFL2與肌纖維組成成分和信號(hào)通路關(guān)鍵成分的相關(guān)性,為建立豬肌纖維性狀的檢測(cè)方法提供前期工作基礎(chǔ)。方法選擇飼養(yǎng)條件相同、同一父系的純種荷包豬、大白豬、長(zhǎng)白豬和杜長(zhǎng)大三元雜交商品豬,按常規(guī)方法分別于1d、30d、60d、90d、120d、150d和180d日齡屠宰母豬各3頭,共計(jì)84頭。屠宰時(shí)測(cè)定肉質(zhì)性狀(肌纖維直徑、剪切力、肉色和p H)。屠宰后背最長(zhǎng)肌的肌肉樣品,采用實(shí)時(shí)定量PCR和Western blot技術(shù)對(duì)肌纖維組成成分(My HCslow、My HC2a、My HC2x、My HC2b、MLC、α-actin)、信號(hào)通路關(guān)鍵成分(MEF2C、sk MLCK、Rho A、AMPKα2、MAPKp38及Ca M)及肌纖維候選基因CFL2的m RNA和蛋白質(zhì)表達(dá)情況進(jìn)行檢測(cè);并對(duì)檢測(cè)數(shù)據(jù)采用SPSS17.0統(tǒng)計(jì)軟件進(jìn)行多重比較方差分析。結(jié)果1、大白和長(zhǎng)白豬的肉質(zhì)性狀變化基本一致。隨著豬的生長(zhǎng)發(fā)育,荷包、大白和長(zhǎng)白豬的肉色顯著加深、剪切力顯著增加、肌纖維直徑顯著增大(P0.05);不同豬種的肉色和p H在120d之后差異不顯著;180d時(shí),各品種豬剪切力差異不顯著,而荷包豬的肌纖維直徑與大白、長(zhǎng)白差異顯著(P0.05)。2、各基因的m RNA檢測(cè)發(fā)現(xiàn):同一基因在不同豬種中有大致相同的生后表達(dá)規(guī)律,但不同品種相同時(shí)間點(diǎn)存在表達(dá)量的顯著差異。大白和長(zhǎng)白豬的My HC總量和肌纖維組分的m RNA表達(dá)量基本一致,而荷包豬60d-150d與大白、長(zhǎng)白豬差異顯著(P0.05);不同品種豬CFL2 m RNA表達(dá)趨勢(shì)基本與My HC總量和肌纖維組分表達(dá)相似。1d-60d不同品種MEF2C m RNA表達(dá)差異不顯著,90d-150d荷包豬顯著高于其他品種,180d荷包豬顯著低于其他品種。sk MLCK m RNA表達(dá)量總體呈上升趨勢(shì);AMPK和MAPKp38 m RNA表達(dá)量卻隨日齡增長(zhǎng)下降。長(zhǎng)白豬30d-180d的Rho A m RNA表達(dá)始終極顯著高于其他豬種(P0.01);荷包豬90d-180d的CAM m RNA表達(dá)都極顯著高于其他品種(P0.01)。3、利用Western Blot方法檢測(cè)蛋白表達(dá),結(jié)果顯示:各品種豬的蛋白表達(dá)趨勢(shì)基本一致,差異在于表達(dá)量不同。荷包豬My HC slow總體極顯著(P0.01)高于其它品種;其他的My HC總量和肌纖維成分(My HC2a、My HC2x、My HC2b、MLC和α-actin),除個(gè)別時(shí)間點(diǎn)差異不顯著,長(zhǎng)白豬總體顯著高于其它品種(P0.05)。荷包豬的CFL2蛋白表達(dá)量自出生至180d始終極顯著高于進(jìn)口豬種(大白、長(zhǎng)白和三元雜交豬)(P0.01)。長(zhǎng)白豬sk MLCK蛋白表達(dá)始終極顯著高于其他品種(P0.01),而荷包豬與大白豬相似且最低。隨豬日齡增長(zhǎng),AMPK表達(dá)量下降,而MAPKp38整體升高。Rho A蛋白,長(zhǎng)白豬1d-150d始終極顯著高于其他品種(P0.01),僅180d低于荷包豬。荷包豬與三元豬的CAM表達(dá)量相似,高于大白和長(zhǎng)白豬。4、相關(guān)分析表明:所有肉質(zhì)性狀均與日齡極顯著相關(guān)(P0.01);肌纖維組成成分中,僅My HC2a與日齡極顯著(P0.01)正相關(guān),α-actin與日齡顯著(P0.05)負(fù)相關(guān);信號(hào)通路AMPKα2和MAPKp38與日齡呈中等程度顯著(P0.05)相關(guān)。My HC2b和My HC2a分別與p H24和肌纖維直徑顯著相關(guān)(P0.05);信號(hào)通路蛋白中AMPKα2與所有肉質(zhì)性狀有關(guān),MAPKp38和Ca M僅與剪切力和肌纖維直徑中、低度相關(guān)。CFL2的表達(dá)僅與My HC slow呈極顯著(P0.01)正相關(guān);My HC總量與MLC也呈極顯著(P0.01)正相關(guān)。除Ca M外,信號(hào)通路關(guān)鍵成分(MEF2C、sk MLCK、AMPKα2、MAPKp38、Rho A)均與肌纖維成分中、低度相關(guān)。結(jié)論1豬的肉質(zhì)性狀和肌球蛋白My HC2a表達(dá)與日齡顯著正相關(guān)。2肌球蛋白My HC2b的含量影響肉質(zhì)的p H,My HC2a的含量影響肌纖維直徑。中國(guó)地方豬—荷包豬肉質(zhì)好于進(jìn)口豬種的原因之一是肌纖維直徑小。3信號(hào)通路AMPKα2與所有肉質(zhì)性狀相關(guān),MAPKp38和Ca M僅與剪切力和肌纖維直徑中低度相關(guān)。4 CFL2可能通過(guò)調(diào)節(jié)My HC slow來(lái)影響肌纖維組成,促進(jìn)肌纖維類型改變進(jìn)而參與影響肉質(zhì)性狀。
[Abstract]:Objective to explore the expression of different varieties of pigs after the birth of muscle fiber composition, key components and related signal pathway genes and interspecific difference, correlation analysis of pork quality traits candidate gene CFL2 and muscle fiber composition key components of signaling pathways and at the same time, provide a basis to establish a detection method of pig muscle fiber traits. The choice of feeding method under the same conditions, the same male purebred Hebao pigs, large white, Landrace and Duroc crossbred pigs three yuan, according to the conventional method respectively in 1D, 30d, 60d, 90d, 120d, 150D and 180d day old mother pig slaughter each 3, a total of 84 head. Determination of meat quality traits at slaughter (diameter of muscle fiber shear force, color and P, H). Slaughter back and the longest muscle samples by real-time quantitative PCR and Western blot technology on muscle fiber composition (My HCslow, My HC2a, My HC2x, My HC2b, MLC, -actin, alpha) signal pathway Components (MEF2C, SK MLCK, Rho A, AMPK MAPKp38 and Ca alpha 2, M) was used to detect the expression of M RNA and protein and muscle fiber candidate gene CFL2; and the test data for multiple comparisons ANOVA using SPSS17.0 statistical software. Results 1 Changes of meat quality traits in large white and Landrace with basically the same. The growth of pigs, large white and Landrace pouch, color significantly deepened, shear stress significantly increased, muscle fiber diameter increased significantly (P0.05); different pig breeds and P H color after 120d 180d, the difference was not significant; the pig breeds shear stress difference is not significant, and Hebao pig muscle fiber the diameter and large white, Landrace was significantly different (P0.05).2, found m RNA to detect the gene, the same gene is roughly the same in different pig breeds after expression pattern, but different varieties at the same time there was significant difference. The amount of large white and Landrace My and total HC M RNA muscle fiber component expression is basically the same, and Hebao pig 60d-150d with large white, Landrace was significantly different (P0.05); different kinds of expression of porcine CFL2 and My HC m RNA the basic trend and total muscle fiber composition similar to different varieties of.1d-60d MEF2C m expression RNA expression had no significant difference, 90d-150d was significantly higher than that of other Hebao pigs 180d varieties, Hebao pigs was significantly lower than the other varieties of.Sk MLCK m RNA expression in the overall upward trend; AMPK MAPKp38 and m RNA expression with age decreased. 30d-180d Rho A m RNA of Landrace was significantly higher than that of the ultimate expression of other breeds (P0.01); the expression of CAM m RNA 90d-180d are significant Hebao pigs higher than other varieties (P0.01).3 expression, using Western method for the detection of Blot protein results showed that all varieties of pig protein expression trend was consistent. The difference lies in the difference in expression of My HC slow overall Hebao pigs (P0.01) were significantly higher In other varieties; the other My HC and total muscle fiber composition (My HC2a, My HC2x, My HC2b, MLC and alpha -actin), in addition to the individual time points were not significantly different, Landrace overall was significantly higher than that of other varieties (P0.05). The expression of CFL2 protein of Hebao pigs from birth to 180d was significantly higher than that of the ultimate import pig breeds (large white, Landrace and three crossbred pigs) (P0.01). The expression of SK MLCK protein was the ultimate Landrace was significantly higher than that of other varieties (P0.01), and the purse and large white pigs. Pigs with similar and minimum age, lower expression of AMPK, MAPKp38 and.Rho increased the overall A protein was 1d-150d, Landrace the ultimate was significantly higher than that of other varieties (P0.01), only 180d is lower than that of Hebao pig Hebao pig and pig three yuan. The expression of CAM was similar to that of large white and Landrace pigs was higher than.4, the correlation analysis showed that: all meat traits were significantly correlated with age (P0.01); muscle fiber components, only My HC2a and age pole Significant positive correlation (P0.01), alpha -actin (P0.05) and age was negatively correlated; moderately significant signal pathway of AMPK alpha 2 and MAPKp38 with age (P0.05) and My HC2b.My correlated HC2a respectively with P H24 and muscle fiber diameter (P0.05); AMPK signaling pathway protein 2 and alpha all MAPKp38 and Ca on meat quality traits, only M and shear force and muscle fiber diameter, low expression of related.CFL2 only with My HC slow (P0.01) showed a significant positive correlation; the total HC and MLC My also showed significant positive correlation (P0.01). In addition to Ca M, a key component of signal pathway (MEF2C SK, MLCK, AMPK, MAPKp38, Rho, alpha 2, A) were related to low and muscle fiber composition, expression. Conclusion 1 pig meat quality traits and myosin My HC2a and age was positively related to.2 My HC2b the effects of the content of myosin P H influence the content of meat, muscle fiber diameter. My HC2a in our local pig - fried pork in good One of the reasons is the kind of pig muscle fiber diameter small.3 signaling pathway AMPK alpha 2 associated with all meat quality traits, MAPKp38 and M only Ca associated with low shear stress and muscle fiber diameter in.4 CFL2 My HC slow may regulate the effect of muscle fibers, promote muscle fiber type and is involved in the change influencing meat quality traits.

【學(xué)位授予單位】：錦州醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S828

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