齐口裂腹鱼和鲤鱼H-FABP基因的克隆及其表达谱

心型脂肪酸结合蛋白（heart fatty acid binding protein, H-FABP）的水平与影响肉质性状的肌内脂肪含量有关,鱼类H-FABP的表达水平对其肌内脂肪含量是否相关仍未见报道.本研究获得齐口裂腹鱼和鲤鱼心脏型脂肪酸结合蛋白基因序列,利用半定量RT-PCR分析其表达特性并测定肌内脂肪含量,比较H-FABP基因在不同生活环境的2种鲤科鱼肌内脂肪沉积中的作用.结果显示,齐口裂腹鱼和鲤鱼H-FABP基因的ORF为402 bp,编码133个氨基酸,它们的氨基酸序列相同,与人、猪、小鼠、斑马鱼、大西洋鲑、虹鳟等的同源性为71.3%~ 90%;H-FABP基因在2种鲤科鱼的心、肌肉、脂肪、肝、脑、脾、肾和鳃等组织中均有表达,肝中的表达量显著高于其它组织（P<0.05）,H-FABP基因的肌肉表达谱在齐口裂腹鱼和鲤鱼中存在明显差异：齐口裂腹鱼中的表达随生长发育呈上升趋势,在大体重鱼（500 g）中的表达显著高于小体重鱼（P<0.05),其表达与肌内脂肪含量呈显著正相关（R=0.370,P<0.05）;H-FABP基因在鲤鱼生长发育中呈下降趋势,而小体重鱼（50~60 g）中的表达显著高于其它大体重鱼(P<0.05),其表达与肌内脂肪含量呈显著负相关（R=-7.083,P<0.01）.据此推测,齐口裂腹鱼和鲤鱼肌肉组织H-FABP基因表达与肌内脂肪关联性的差异可能与2种鱼的生活环境不同有关.     

猪H-FABP基因多态性与肌内脂肪含量的关联分析

以山西白猪为实验材料,采用索氏脂肪抽提法对其背最长肌的肌内脂肪含量进行了测定。根据心脏脂肪酸结合蛋白(heart fatty acid-binding protein,H-FABP)基因的PCR-RFLP多态性,应用SPSS软件,用最小二乘法拟合一般线性模型(GLM)分析了H-FABP基因的不同位点的基因型对肌内脂肪含量的影响。结果表明,基因型hh、dd、BB对肌内脂肪含量影响的效应值最大。证明H-FABP基因可以作为肌内脂肪含量的侯选基因。     

中国美利奴羊H-FABP基因多态性及其与部分肉质性状相关性的研究

利用聚合酶链式反应-单链构象多态（PCR-SSCP）技术检测中国美利奴羊(Chinese Merino)心脏型脂肪酸结合蛋白基因（H-FABP）外显子2的单核苷酸多态性（SNPs）和遗传多态性,分析其与肌内脂肪（IMF）含量、肌纤维直径和肌纤维密度的相互关系,为该品种绵羊的分子标记辅助选择提供理论依据。结果显示,H-FABP基因外显子2有AA、AB和BB 3种基因型,AA型和BB型在778位均发生了C缺失,939位均发生了A→G转换,BB型还在789位发生了T→C转换,该突变导致所编码氨基酸发生了缬氨酸→丙氨酸的替换;BB型为IMF的优势基因型,与AB型相比差异显著(P<0.05),与AA型相比差异极显著（P<0.01);BB型对肌纤维直径存在负相关。结果提示,中国美利奴羊H-FABP基因外显子2具有多态性,该基因可能是中国美利奴羊肉质性状的主效基因,或者与控制这些性状的主效基因相连锁。     

MyoG基因的遗传效应分析

实验采用PCR-RFLP技术分析了不同品种猪MyoG基因3′端MspⅠ位点的多态性,应用单标记回归模型分析了不同基因型与相关性状间的关联性及不同等位基因的遗传效应。结果表明：N等位基因能极显著地增加胴体瘦肉率和眼肌面积,降低皮脂含量（P<0.01）,改善胴体产肉量和提高胴体品质;同时,不同基因型对肉质性状的遗传影响作用较大,表现为N等位基因能极显著地降低猪肉品质,使pH值、肉色和肌内脂肪含量降低,并使肌肉的系水力变差（P<0.01）。N等位基因对增加胴体瘦肉率的加性效应值和显性效应值分别为3.929%和-0.602%;对增加眼肌面积的加性效应值和显性效应值分别为2.0985 cm2和-0.5775 cm2;对皮脂率的加性效应值为-3.0245%,显性效应值为-0.4045%。N等位基因对pH1的加性效应值和显性效应值分别为-0.167和0.034;对贮藏损失的加性效应值和显性效应值分别为0.558和-0.347;对肌内脂肪含量的加性效应值和显性效应值分别为-0.963和-0.217。但MyoG基因3′端MspⅠ位点的突变对FOM肉脂仪测定的胴体等级性状的影响不显著（P>0.05）。     

猪FUT1基因对肉质和胴体性状的影响

测定139头杂交猪(大白猪和梅山猪)的14个肉质性状和8个胴体性状,用PCR-RFLP方法检测FUT1基因型。分析猪FUT1基因型间肉质和胴体性状差异,发现AA基因型猪3个部位肌肉pH值均比AG基因型的高,其中pH(LD)达到显著水平(P<0.05)。AA基因型猪肌肉系水力显著高于AG猪的系水力(91.02% VS 86.70%,P<0.05)。AA基因型猪的肉色值显著高于AG猪的(P<0.05)。AA基因型猪三个部位肌肉膘厚值均较低,其中最后肋骨膘厚和倒数三四肋骨膘厚分别比AG基因型猪的低4.26 mm和3.96 mm(P<0.05)。AA基因型猪瘦肉率比AG基因型猪的高3.31% (53.46% VS 50.15%,P<0.05)。以上结果表明FUT1基因的AA基因型对肉质和和胴体性状具有显著的正遗传效应,这对于在抗病育种中应用该基因十分有利。     

西部地区主要猪种H-FABP基因多态性,IMF含量及不同基因型脂肪细胞脂滴量的关系

H-FABP是FABP家族成员之一,在长链脂肪酸的吸收和代谢平衡中发挥关键作用,但它对猪IMF含量的影响还知之甚少,对中国西部地区的猪种更是如此.文章利用PCR-RFLP（HinfⅠ、HaeⅢ和Msp Ⅰ 3种限制性内切酶）分子标记技术,检测了杜洛克猪、长白猪、大白猪、内江猪、荣昌猪、汉江黑猪、汉中白猪、八眉猪和野猪共计265头猪H-FABP基因5＇-上游区和第二内含子区的遗传变异,利用最小二乘模型分析了H-FABP基因对猪肌内脂肪含量的遗传效应,并运用猪脂肪细胞培养,油红O染色和TG测定等技术检测了H-FABP基因不同基因型脂肪细胞内脂滴的形态和沉积的量.结果表明：（1）在HinfⅠ-RFLP位点上,上述品种和野猪均存在多态性,其中大白猪、八眉猪、汉江黑猪、汉中白猪和野猪表现为低度多态,杜洛克、长白猪、内江猪和荣昌猪为中度多态;除汉江黑猪（P＜0.05）和野猪（P＜0.01）外,其他猪种基因频率和基因型频率都处于Hardy-Weinderg平衡状态（P＞0.05）;而在HaeⅢ-RFLP和Msp Ⅰ-RFLP位点上,仅内江猪、荣昌猪、汉江黑猪和八眉猪为单态;（2）9种基因型对肌内脂肪（IMF）含量的影响,HH＞Hh＞hh,DD＜Dd＜dd,AA＜Aa＜aa,遗传效应值分别为：3.89,3.42,3.17,2.27,2.49,2.91,2.28,2.70,2.95,H-FABP基因可显著地提高IMF含量（P＜0.05）;（3）aaddHH型的脂肪细胞脂滴的形态,密度和含量与其他基因型细胞差异显著（P＜0.05）.结果提示：可通过提高＂aaddHH＂基因型的频率来增加IMF含量,达到改善猪肉质的目的.     

脂肪分化相关蛋白基因多态性与优质鸡屠宰性状和肌内脂肪含量的相关性研究

对脂肪分化相关蛋白(Adipocyte Differentiation-Related Protein, ADFP)基因的外显子进行SNPs 检测, 探讨其作为鸡脂肪性状候选基因的可能性。实验以四川省畜牧科学研究院和大恒家禽育种有限公司培育的优质肉鸡新品系为素材, 采用PCR-SSCP的方法进行SNPs 检测和基因型的分析。结果找到3个SNPs位点: 4 079位由A→T(位点A)、4 843位由C→T(位点B)和7 070位由A→G(位点C)。单位点基因型对屠宰性状的遗传效应分析表明, 位点A的基因型对腿肌率、腹脂重、腹脂率和肌内脂肪含量有显著性影响(P < 0.05), 位点B的基因型对活重和屠体重均有显著性影响(P < 0.05), 位点C的基因型对胸肌重和肌内脂肪含量有显著性影响(P < 0.05), 对胸肌率有极显著性影响(P < 0.01)。初步推断ADFP基因可能是影响鸡脂肪性状的主效基因或与主效基因连锁, 推测可以利用多态位点A和C对鸡腹脂重、腹脂率和肌内脂肪含量进行标记辅助选择。     

西部地区主要猪种和野猪H-FABP基因分子标记

利用PCR-RFLP(Hinf I、Hae Ⅲ和Msp I 3种限制性内切酶)分子标记技术,检测了杜洛克猪、长白猪、大白猪、内江猪、荣昌猪、汉江黑猪、汉中白猪、八眉猪和野猪共计265头猪H-FABP基因5′-上游区和第二内含子区的遗传变异,并利用最小二乘模型分析了H-FABP基因对猪肌内脂肪含量的遗传效应。结果表明：（1）在Hinf I-RFLP位点上,上述品种和野猪均存在多态性,其中大白猪、八眉猪、汉江黑猪、汉中白猪和野猪表现为低度多态,杜洛克、长白猪、内江猪和荣昌猪为中度多态;除汉江黑猪（P<0.05）和野猪（P<0.01）外,其他猪种基因频率和基因型频率都处于Hardy-Weinderg平衡状态（P>0.05）;而在Hae Ⅲ-RFLP和Msp I-RFLP位点上,仅内江猪、荣昌猪、汉江黑猪和八眉猪为单态;（2）9种基因型对肌内脂肪（IMF）含量的影响,HH>Hh>hh,DD相似文献   

H-FABP基因多态性与江苏地方山羊品种IMF含量的关系

目的:探讨H -FABP基因多态性与江苏地方山羊品种肌内脂肪含量之间的相关性.方法:PCR - SSCP标记技术检测2个江苏地方山羊品种H -FABP基因部分外显子和内含子区域的多态性,利用最小二乘法分析其与IMF含量的关系.结果:在H - FABP基因第2外显子区域发现1个多态性位点,分别定义为GG和GC2种基因型.构建的固定效应模型显示,基因型GC的IMF含量含量高于基因型GG,在背最长肌和腿肌中GC基因型个体的IMF含量显著高于GG基因型(P＜0.05).结论:H-FABP基因第2外显子132 bp位点处GC基因型具有使IMF含量提高的遗传效应.     

鸡H-FABP基因多态性及其与屠宰性能的关联分析

以90日龄草科鸡为实验材料, 对H-FABP基因进行PCR扩增, 采用PCR-SSCP技术结合测序分析了H-FABP基因在草科鸡中的多态性。结果表明: 草科鸡中存在332 G→A、534 G→A、835 C→T、1131→A、1294C→A、2329C→T、2372C→T、2636C→T等8处突变。经过基因型与屠宰性能的关联分析得知: 在7对产生多态的引物中, 只有第1对引物的各基因型与活重、屠体重、胸肌重、腿肌重和半净膛重差异显著(P<0.05); 全净膛重在不同基因型间差异极显著(P<0.01)。 由此推测, H-FABP可能对于鸡屠宰性能具有很大的影响或与控制屠宰性能的主基因连锁。     

A study of associations of the H-FABP genotypes with fat and meat production of pigs

Although the content of intramuscular fat (IMF) influences significantly meat quality, it can be estimated only after the slaughter of animals. Variants of the H-FABP gene were suggested as candidate genes influencing the variability of IMF. The effect of H-FABP - HinfI polymorphism on the content of IMF, backfat thickness, the weight and percentages of major meat parts and of the leg in carcass weight was studied in a group of 97 pigs (46 gilts and 51 barrows) of Large White and Landrace breeds using the test of fattening capacity and carcass value. In the set of experimental animals, the frequencies of genes were H = 0.75 +/- 0.03 and h = 0.25 +/- 0.03. Biometric analyses did not corroborate differences among different H-FABP - HinfI genotypes and all the traits under study. Only in genotypes HH and Hh the differences between least-square means of phenotypic IMF values under study were close to the limit of significance (P = 0.06).     

Test for positional candidate genes for body composition on pig chromosome 6

One QTL affecting backfat thickness (BF), intramuscular fat content (IMF) and eye muscle area (MA) was previously localized on porcine chromosome 6 in an F₂ cross between Iberian and Landrace pigs. This work was done to study the effect of two positional candidate genes on these traits: H-FABP and LEPR genes. The QTL mapping analysis was repeated with a regression method using genotypes for seven microsatellites and two PCR-RFLPs in the H-FABP and LEPR genes. H-FABP and LEPR genes were located at 85.4 and 107 cM respectively, by linkage analysis. The effects of the candidate gene polymorphisms were analyzed in two ways. When an animal model was fitted, both genes showed significant effects on fatness traits, the H-FABP polymorphism showed significant effects on IMF and MA, and the LEPR polymorphism on BF and IMF. But when the candidate gene effect was included in a QTL regression analysis these associations were not observed, suggesting that they must not be the causal mutations responsible for the effects found. Differences in the results of both analyses showed the inadequacy of the animal model approach for the evaluation of positional candidate genes in populations with linkage disequilibrium, when the probabilities of the parental origin of the QTL alleles are not included in the model.     

H-FABP、MC4R、ADD1基因多态性在3个猪群中分布及其与肌内脂肪和背膘的相关研究

具有不同遗传特性的猪种具有不同的肉质性状。尤其是地方品种和引进品种间在肉质性状存在极大的差异。在已有的研究中H-FABP,MC4R,ADD1基因同肌内脂肪或背膘相关。利用梅山猪、苏太猪和杜×长×大猪为试验动物,研究上述3个基因的多态性分布和多态性同肌内脂肪和背膘的相关性。结果表明：3个基因的多态性分布在不同猪种间存在极显著的差异,这种差异可能是肌内脂肪（IMF）或背膘（BF）不同的主要原因之一。连锁分析表明：H-FABP和ADD1基因多态性同IMF有显著的相关,但是同BF没有显著的相关;MC4R基因的多态性同IMF和BF都有显著相关性。说明：H-FABP和ADD1基因多态性有可能应用到提高IMF,同时不影响BF的育种实践中。      

Effects of feeding and rearing systems on growth, carcass composition and meat quality in pigs

Animal growth performance and quality of pork depend on the interactive effects of pig genotype, rearing conditions, pre-slaughter handling, and carcass and meat processing. This paper focuses on the effects of feeding and rearing systems (feeding level and diet composition, housing, production system, etc.) on growth performance, carcass composition, and eating and technological qualities of pork. The feeding level and protein : energy ratio can be used to manipulate growth rate or composition of weight gain. Restricted feed allowance strongly reduces growth rate and carcass fatness and also intramuscular fat (IMF) level, resulting in decreased meat tenderness or juiciness. Expression of compensatory growth due to restricted followed by ad libitum feeding modifies the composition of weight gain at both carcass and muscle levels, and may improve meat tenderness due to higher in vivo protein turnover. Decreasing the protein : energy ratio of the diet actually increases IMF and improves eating quality, but gives fatter carcasses. In contrast, a progressive reduction in the protein : energy ratio leads to similar carcass composition at slaughter but with higher IMF. Technological meat traits (pH1, pHu, colour, drip loss) are generally not affected by the level or protein : energy in feed. Modification of fatty acid composition and antioxidant level in meat can be obtained through diet supplementations (e.g. vegetable sources with high n-3 fatty acids), thereby improving the nutritional quality of pork. Influences of pig rearing system on animal performance, carcass and meat traits result from interactive effects of housing (floor type, space allowance, ambient temperature, physical activity), feeding level and genotype in specific production systems. Indoor enrichment (more space, straw bedding) generally increases growth rate and carcass fatness, and may improve meat juiciness or flavour through higher IMF. Outdoor rearing and organic production system have various effects on growth rate and carcass fatness, depending on climatic conditions and feed allowance. Influence on meat quality is also controversial: higher drip and lower pHu and tenderness have been reported, whereas some studies show improved meat juiciness with outdoor rearing. Discrepancies are likely due to differences between studies in rearing conditions and physiological responses of pigs to pre-slaughter handling. Specific production systems of the Mediterranean area based on local breeds (low growth rate, high adiposity) and free-range finishing (pasture, forests), which allows pig to express their genetic potential for IMF deposition, clearly demonstrate the positive effects of genotype × rearing system interactions on the quality of pork and pork products.     

Detection of differentially expressed genes in the longissimus dorsi of Northeastern Indigenous and Large White pigs

Recent attention in pig breeding programs has focused on the improvement of pork quality in response to increasing consumer demands. Compared to the fatty-type Northeastern Indigenous (Chinese) breed of pigs, the lean-type Large White has lower intramuscular fat and inferior eating quality from the perspective of the Chinese consumer. In order to investigate the molecular basis of differences in pork quality in Chinese indigenous and Western breeds, longissimus dorsi samples were collected from three adult Northeastern Indigenous and three adult Large White pigs. The RNAs were extracted and hybridized to the porcine Affymetrix GeneChip. Microarray analysis demonstrated differential expression of 1134 genes of which 401 have a known function. One hundred and thirty-six genes were up-regulated and 998 down-regulated in Northeastern Indigenous breed compared to Large White pigs. We screened 10 genes as candidate genes associated with pork quality. We investigated a single nucleotide polymorphism in the 5' regulatory region of the gene FABP4 in 65 Songliao black swine, using PCR-single-strand conformational polymorphism. We found this polymorphism to be highly significantly associated with marbling and intra-muscular fat content (P ≤ 0.01). Genotype BB had higher marbling than AB and AA, but there was no significant difference between AB and AA. Genotype BB and AB had higher intra-muscular fat content than AA, but there was no significant difference between BB and AB. These results help to elucidate the genetic mechanisms behind differences in pork quality and provide a theoretical basis for selection and genetic improvement of meat quality traits in pigs.     

The rearing system modulates biochemical and histological differences in loin and ham muscles between Basque and Large White pigs

Conventional pork production, based on highly selected breeds for growth efficiency and carcass leanness, is generally considered to decrease pork quality. In contrast, non-selected breeds produced in extensive systems are associated with high pork quality, which is generally attributed to higher intramuscular fat (IMF) content and less glycolytic muscle metabolism. The present study aimed to determine biochemical, histological and quality traits of loin and ham muscles of pigs from selected Large White (LW) and local French, non-selected Basque (B) breeds. Pigs were reared in a conventional indoor (C, slatted floor), alternative (A, indoor bedding and outdoor area) or extensive system (E, free range, B pigs only). A total of 100 castrated males were produced in 2 replicates, each containing 5 groups of 10 pigs based on breed and system: LWC, LWA, BC, BA and BE. The glycolytic longissimus muscle (LM) and semimembranosus muscle (SM), and the deep red (RSTM) and superficial white (WSTM) portions of semitendinosus muscle (STM) were studied at 145 kg BW. Overall, breed induced stronger effects on muscle traits than the rearing system, among which the E system induced greater changes. The lower muscle growth of B pigs was associated with fewer muscle fibers and a smaller cross-sectional area (CSA) of glycolytic fibers (P < 0.01). The SM was less glycolytic and more oxidative in B than in LW pigs (P < 0.001). The WSTM followed a similar trend, with a larger relative area of type I fibers in B pigs. In contrast, the LM and RSTM were more oxidative in LW pigs. B pigs had higher IMF content and ultimate pH in all muscles, along with lower glycolytic potential, less light and redder meat in the LM and SM (P < 0.001). Compared to the C system, the A system induced only a shift towards a more oxidative metabolism in the LM and a smaller fiber CSA in the RSTM of LW pigs (P < 0.05), without influencing pork quality traits. Compared to BC pigs, BE pigs had a more oxidative and less glycolytic muscle metabolism, along with higher ultimate pH, lower lightness and redder meat (P < 0.01), but similar IMF content. Overall, results indicate that influences of breed and rearing system on muscle properties depend on muscle type, and that IMF content and fiber-type composition are unrelated traits that can be modified independently by genetic or rearing factors.