哈萨克马和焉耆马不同组织中IGFBP-5基因表达差异研究

试验旨在探求胰岛素样生长因子结合蛋白-5（insulin-like growth factor binding protein-5,IGFBP-5）基因在哈萨克马和焉耆马不同组织部位中的mRNA表达规律。采用实时荧光定量PCR技术检测在哈萨克马和焉耆马心脏、肝脏、脾脏、肺脏、肾脏、小肠、大肠、盲肠、肋间肌、背最长肌、臂肌和臀肌等不同组织中IGFBP-5基因mRNA表达量,同时比较该基因在哈萨克马和焉耆马相同组织中的表达差异。结果表明,哈萨克马和焉耆马表达量最高的均为背最长肌,其次是臂肌,并显著高于心脏、肝脏、脾脏、肺脏、肾脏、小肠、大肠、盲肠等内脏组织表达量（P<0.05）,大肠中的表达量最低;焉耆马的肾脏、小肠、大肠、盲肠、背最长肌和臂肌中表达量高于哈萨克马相同部位的表达量,其中焉耆马背最长肌和大肠中的表达量极显著高于哈萨克马的相同部位（P<0.01）,小肠和盲肠中的表达量显著高于哈萨克马的相同部位（P<0.05）。本试验为深入研究IGFBP-5基因的生物学功能,以及对中国马生产性能的遗传改良提供理论依据。     

Electromyographic activity of the longissimus dorsi muscles in horses when walking on a treadmill

The pattern of electromyographic activity of the equine long back muscle at the walk has not yet been reported. The aim of this study was to use surface electromyography to measure activity of the longissimus dorsi muscles of horses walking on a treadmill. Fifteen horses without back pain were used and electromyographs were recorded bilaterally from the longissimus dorsi muscles at the level of T12, T16 and L3. Mean electromyograph activity and mean motion were calculated for each horse. At the walk, only one maximum activity for each longissimus dorsi muscle was detected during each motion cycle and this was highest at T12 and lowest at L3. Activity of the longissimus dorsi muscles at the walk is mainly responsible for stabilisation of the vertebral column against dynamic forces. At T12 the high maximum activity could contribute to the development of muscle pain at this site.     

普氏野马与两种新疆地方品种马的ELA-DRA*exon2的多态性分析

为了解普氏野马、哈萨克马与焉耆马的ELA-DRA*exon2的多态性以及等位基因频率分布。本研究采用PCR-SSCP技术对普氏野马、哈萨克马与焉耆马的ELA-DRA*exon2进行分型,并对不同等位基因进行核苷酸与氨基酸分析。结果显示:127匹马中共出现7种基因型,3种纯合子分别记为AA、BB、CC;4种杂合子分别记为AB、AC、BC、AD。AA基因型为哈萨克马与焉耆马的优势基因型,普氏野马以CC基因型为主。经χ2检验后,3种类型马的等位基因频率和基因型频率均处于Hardy-Weinberg平衡状态。PIC值与He值分析表明,3种类型马均属于中度多态,且普氏野马的PIC值与He值比哈萨克马与焉耆马低。     

Research on IgG Fc Receptor Gene Expression in Different Parts of the Intestine of Yili Horse

The research was designed to analyze the rule of mRNA expression of IgG Fc receptor(FcR) in intestine from Yili horse aged at two-years-old by RT-PCR method, which aimed at providing data for researches on immunology mechanism and digestive physiology of equine, improving immunology function of horses, enhancing of nutrients absorbing, decreasing mortality.The results showed that the expression of IgG FcR gene in small gut was significantly higher than that of cecum (P< 0.05);it was also higher than that of large gut, its expression in large gut was higher than that of cecum(P> 0.05).The expression of IgG FcR had been identified in intestine of horses at two-year-old, it was different of immunology mechanism among horses, human and rat;Moreover, the expresson of IgG FcR in small gut was the highest among intestine, its expression rule was similar as other mammal animals (piggy and sheep).Above all, not only could be small gut thought as a digestive and absorption organ in nutrients, but also a important immunology organ in terms of its function, each plays special role in physiology of animal body.     

胎儿期与成年期蒙古马骨骼肌肌纤维类型转化研究

为探索影响蒙古马胎儿期和成年期肌纤维类型差异机理.本研究选取3匹4月龄胎儿(两母一公)与3匹5岁健康成年母马身体4块分布全身、具有代表性的肌肉组织(长臂三头肌、夹肌、背最长肌、臀中肌)作为一个整体.胎儿期蒙古马肌纤维和成年期蒙古马肌纤维因存在差异各做为一组,试验进行3个生物学重复.首先对蒙古马骨骼肌肌肉样品进行免疫组化...     

The Expression Characteristics of MYNN Gene in Different Tissues and Muscles of Pig

This study was aimed to investigate the expression characteristics of myoneurin (MYNN) gene in different tissues and its developmental expression in muscles (longissimus dorsi, biceps femoris and psoas major), cerebellum, liver, pancreas, kidney, stomach, spleen and lung tissues of pigs. The expression characteristics of MYNN mRNA in 11 different tissues including heart, liver, spleen, lung, kidney, cerebellum, small intestine, pancreas, stomach, biceps femoris and fat of Large White pigs and Mashen pigs at the age of 90 days and the developmental expression patterns in muscle, cerebellum, liver, pancreas, kidney, stomach, spleen at three strages (1, 90, 180 days) of Large White pigs and Mashen pigs were studied by Real-time PCR. The results showed that MYNN was widely expressed in various tissues of pigs, and there was significant difference among the tissues(P < 0.05; P < 0.01); The expression of MYNN in muscle, liver, pancreas, cerebellum, kidney, stomach, spleen, lung tissues were significant difference at three development stages of Large White pigs and Mashen pigs(P < 0.05; P < 0.01), and also had a specific rule, which indicated that it may play an important role in these pig tissues. The expression of MYNN gene could related to the tissue, age and the genetic background of breeds. The results of this study provided a better understanding of the biological functions of pig MYNN. Further studies are required to determine its molecular mechanisms, especially in the regulation of skeletal muscle development.     

MYNN基因在猪不同组织及肌肉中的表达规律

试验旨在研究myoneurin（MYNN）基因在猪不同组织中的表达特征及其在肌肉（背最长肌、股二头肌和腰大肌）、小脑、肝脏、胰脏、肾脏、胃、脾脏、肺脏组织中的发育性表达规律。采用实时荧光定量PCR技术研究猪MYNN mRNA在90日龄大白猪和马身猪的心脏、肝脏、脾脏、肺脏、肾脏、小脑、小肠、胰脏、胃、股二头肌及脂肪共11个组织中的表达谱,以及在大白猪和马身猪1、90、180日龄3个发育阶段的肌肉、小脑、肝脏、胰脏、肾脏、胃、脾脏、肺脏组织中的发育性表达规律。结果表明,MYNN在猪的各种组织中广泛表达,且各组织间表达差异显著或极显著（P < 0.05;P < 0.01）;MYNN在大白猪和马身猪的肌肉、小脑、肝脏、胰脏、肾脏、胃、脾脏、肺脏组织中的不同发育阶段表达差异显著或极显著（P < 0.05;P < 0.01）,并具有特定规律,由此推测其可能在猪的这几种组织中发挥重要作用。MYNN基因的表达与组织、日龄及品种的遗传背景有关。本试验为研究猪MYNN基因的生物学功能提供了依据,但还需要深入的研究来探索其作用的具体机制,尤其是在骨骼肌发育中的调节机制。     

草原红牛与天一冈山黑牛背最长肌IGFBP-1、IGFBP-2、IGFBP-3、IG-FBP-4及IGFBP-5基因表达差异

本研究采用实时荧光定量RT-PCR技术检测了IGFBP-1、IGFBP-2、IGFBP-3、IGFBP-4及IGFBP-5基因在草原红牛与天一冈山黑牛背最长肌中的表达情况,旨在探求这些基因在草原红牛与天一冈山黑牛背最长肌中的表达规律。结果表明,所有5个基因在草原红牛与天一冈山黑牛背最长肌中均有表达;同时,IGFBP-1基因的mRNA在2种牛背最长肌中的相对表达量存在极显著的差异（P〈0.01）,其在草原红牛中的相对表达量是在天一冈山黑牛中的1.42倍;IGFBP-2基因的mRNA在2种牛背最长肌中的相对表达量存在显著的差异（P〈0.05）,其在草原红牛中的相对表达量是在天一冈山黑牛中的3.55倍;其他3个基因的相对表达量没有显著差异。本研究为深入研究IGFBP-1、IGFBP-2、IGFBP-3、IGFBP-4和IGFBP-5基因的生物学功能、了解这些基因对肉质性状的影响,以及对我国肉牛的改良提供理论依据。     

猪PPARs基因组织表达特性的研究

以成年母猪为研究材料,采用RT-PCR半定量法对猪PPARα、β/δ和γ基因组织表达特点进行了研究。结果表明:在检测的18种组织中除胰腺组织外,3种PPAR亚型在其他17种组织中均有表达。表达量高低依次为PPARα,子宫绒毛膜>皮下脂肪>卵巢>大肠>脾脏>大脑>肾上腺>心脏>肺>小肠>脊髓>子宫蜕膜>胃>肝脏>背最长肌>膀胱>肾脏;PPARδ,子宫绒毛膜>卵巢>大肠>脾脏>肝脏>胃>皮下脂肪>大脑>肺>肾上腺>子宫蜕膜>脊髓>背最长肌>心脏>肾脏>小肠>膀胱;PPARγ,背最长肌>皮下脂肪>卵巢>脾脏>肺>大肠>膀胱>子宫绒毛膜>子宫蜕膜>心脏>胃>肝脏>肾脏>大脑>脊髓>肾上腺>小肠。3种亚型PPAR在卵巢和/或子宫绒毛膜中都有较高的表达,提示它们与猪的繁殖性能相关。     

Pax7基因在猪不同组织中的表达特征及其在背最长肌中的发育性表达规律研究

本试验旨在探究Pax7(paired box 7)基因在猪不同组织中的表达特征及其在背最长肌中的发育性表达规律.采用实时荧光定量PCR(qRT-PCR)技术和Western blotting技术检测了Pax7基因在1日龄马身猪心脏、肝脏、脾脏、肺脏、肾脏、胃、小肠、下丘脑、小脑、背最长肌、腰大肌、股二头肌12种组织中的mRNA和蛋白质表达谱,以及马身猪和大白猪从出生1日龄到180日龄7个发育阶段(1、30、60、90、120、150、180日龄)背最长肌中的发育性表达规律.结果表明,Pax7基因mRNA在背最长肌、腰大肌、股二头肌、下丘脑和小脑组织中表达,而Pax7蛋白仅在背最长肌、腰大肌和股二头肌中表达.背最长肌中Pax7基因mRNA和蛋白质的发育性表达趋势在马身猪和大白猪中基本一致.在mRNA水平上,30日龄的表达量最高,极显著高于其他日龄(P <0.01);1日龄的表达量次之;其他日龄维持低表达的稳定状态.Pax7蛋白表达量在1日龄时最高,极显著高于其他日龄(P <0.01);30日龄次之;其他日龄维持低表达的稳定状态.从出生1日龄到180日龄,马身猪背最长肌中Pax7蛋白的表达量均显著或极显著地高于大白猪(P <0.05;P <0.01).Pax7 基因的表达与组织、日龄及品种的遗传背景有关.     

猪A-FABP基因真核表达载体的构建及其组织表达

本实验以长白猪背最长肌为实验材料,进行组织RNA的提取及A-FABP基因的克隆,构建p EGFPN1-A-FABP真核表达载体,并通过脂质体转染成纤维细胞,48 h观察荧光表达。同时应用荧光定量PCR法检测猪7种不同组织(心脏、肝脏、脾脏、肺脏、肾脏、背最长肌、腿肌)中A-FABP基因的差异表达。结果表明:成功构建p EGFP-N1-A-FABP融合表达载体,并在细胞中表达绿色荧光蛋白;A-FABP基因在7种组织中均有表达,其中背最长肌和腿肌中A-FABP基因的表达量最高,与其他组织相比差异极显著(P0.01),而脾脏和肾脏中的表达量相对于心脏、肝脏和肺脏差异显著(P0.05),而A-FABP基因表达量在心脏、肝脏和肺脏中差异不显著(P0.05)。表明该基因能够在真核载体和细胞中表达,并且在猪不同组织的表达具有差异性。     

牛多个组织microRNASolexa测序与生物信息学分析

本研究旨在发掘牛更多的microRNA(miRNA)信息及新的miRNA序列,为进一步研究miRNA的生物学功能奠定理论基础.运用高通量测序技术对来自一头西门塔尔公牛和一头荷斯坦母牛的多个组织的小RNA进行混池测序,对测序结果进行了生物信息学分析.随机选取了一条成熟miRNA:bta-miR-2346-5p和两条新预测的miRNAs:novel_miR-48和novel_miR-86,采用茎环RT-PCR进行验证.共鉴定了604条miRNAs,其中429条为牛的已知miRNAs,175条为新预测的miRNAs.通过茎环RT-PCR发现选取的这3条miRNAs在背最长肌、心肌、肝脏、脾脏、肺脏、肾脏、大肠、小肠中均有表达,证明了高通量测序结果的准确性.本试验得到的数据在一定程度上丰富了牛miRNA的数量和种类,为后续牛或其他物种的miRNA相关生物学研究提供了更加详实的信息.     

Study on the Expression Profile of Pax7 Gene and its Developmental Expression in Longissimus Dorsi of Pigs

The aim of this study was to investigate the expression profile of Pax7 (paired box 7) gene in different tissues and its developmental expression in longissimus dorsi of pigs.The expression profiles of Pax7 gene at the levels of mRNA and protein in 12 different tissues including heart,liver,spleen,lung,kidney,stomach,small intestine,hypothalamus,cerebellum,longissimus dorsi,biceeps femoris,psoas major of Mashen pigs at birth and the developmental expression patterns in longissimus dorsi at seven stages (1,30,60,90,120,150,and 180-day old) of Mashen and Large White pigs were studied by quantitative Real-time PCR and Western blotting.The results showed that the Pax7 mRNA was expressed in longissimus dorsi,biceeps femoris,psoas major,hypothalamus,and cerebellum,wherease the Pax7 protein was only expressed in skeletal muscles of longissimus dorsi,biceeps femoris,and psoas major.The developmental expression patterns of Pax7 mRNA and protein in Mashen pig was basically in accordance with those in Large White pig.The mRNA expression amount of Pax7 gene in longissimus dorsi was the greatest at 30-day old (P <0.01),followed by the expression at 1-day old,and kept lower stable levels at other stages both in Mashen and Large White pigs.At the stages of 30-day,90-day,and 180-day old,Pax7 mRNA expression amounts in Mashen pig were significantly higher than those in Large White pig (P <0.01).The Pax7 protein expression at 1-day old was the greatest (P <0.01),followed by it at 30-day old,which were significantly greater than those at other stages,at which Pax7 expression was in lower stable levels.The Pax7 protein expression from one-day old to 180-day old in Mashen pig was significantly greater than those in Large White pig (P <0.05;P <0.01).The expression of Pax7 gene could associated with tissue,age,and background of pig breed.     

关岭牛肌球蛋白重链1基因5'侧翼启动子的克隆和生物信息学分析

本试验旨在进行关岭牛肌球蛋白重链1（myosin heavy chain 1,MYH1）5'侧翼启动子的克隆和生物信息学分析。采集关岭牛血液及组织样品（背最长肌、后腿肌、心脏、肝脏、小肠、脂肪组织）,利用PCR方法扩增关岭牛MYH1基因5'侧翼区,构建关岭牛pUCm-T-MYH1克隆载体,并对MYH1基因5'侧翼区进行生物信息学分析,最后利用实时荧光定量PCR法测定了MYH1基因在关岭牛不同组织中的表达。结果显示,本试验成功获得1 373 bp（-1 360~+12 bp）的MYH1基因5'侧翼启动子序列;利用生物信息软件对获得的克隆序列进行分析发现,MYH1基因5'侧翼启动子序列存在5处可能的转录起始点和多个潜在转录因子结合位点;关岭牛与金丝猴、野猪、家鼠、藏羚羊、野驴的MYH1基因5'侧翼区保守性较强的区域为转录起始点上游-400~+100 bp,推测转录起始点上游-400~+75 bp可能是其核心启动子区域。实时荧光定量PCR分析发现,MYH1基因在关岭牛背最长肌和后腿肌中高表达,在心脏、肝脏、小肠、脂肪组织中表达量很低,说明MYH1基因表达具有组织特异性。     

马身猪和大白猪不同组织DECR1基因的表达

旨在研究DECR1基因在猪不同组织和品种中mRNA和蛋白水平的表达规律,探讨该基因与脂肪代谢的关系。以山西马身猪与大白猪为试验材料,提取肝脏、心脏、肾脏、脾脏、肺脏、胃、小肠、皮下脂肪和背最长肌组织的总RNA和总蛋白,应用实时荧光定量PCR技术检测DECR1基因在2个品种各组织中mRNA的相对表达量,采用Western blot技术对各组织中DECR1蛋白进行半定量分析。实时荧光定量PCR结果显示:DECR1基因在各组织中均有表达,组织之间的表达量存在极显著差异(P<0.01),DECR1基因在肝脏、皮下脂肪与心脏中为高丰度表达;在不同品种的皮下脂肪组织中,大白猪DECR1的mRNA表达极显著高于马身猪(P<0.01)。Western blot检测结果显示:DECR1在各组织中均有表达,不同组织的表达量存在极显著差异(P<0.01),在皮下脂肪、肝脏与小肠中高表达;不同品种的皮下脂肪组织中,大白猪DECR1蛋白的表达显著高于马身猪(P<0.05)。猪DECR1基因在不同组织的表达差异可能与脂肪代谢和脂肪沉积有关。     

Study on Melanocortin-4 Receptor mRNA Expression in Goats Grazed in Leigongshan Mountain Area

In order to understand the differential expression among tissues or breeds of grazed goat,β-actin gene was used as control,the expression levels of melanocortin-4 receptor (MC4R) mRNA in 7 tissues of Qiandongnan small Xiang goat,Guizhou White goat,Guizhou Black goat,Qianbei Ma goat and Nanjiang Yellow goat were detected by Real-time fluorescence quantitative PCR technique with double standard curve method.The results showed that MC4R mRNA was expressed in all 7 tissues of 5 grazed goat breeds,the expression levels of subcutaneous fat and kidney were the highest,the expression levels of liver was the second,the expression levels of lung and heart were the third,and the expression levels of semimembranosus muscle and longissimus muscle were the lowest.There were no differences of the expression level of MC4R mRNA in liver among different breeds.In subcutaneous fat,kidney,lung,heart,semimembranosus muscle and longissimus muscle,the highest expression levels of MC4R mRNA were all observed in Guizhou White goat.The results suggested that for grazed goats,the expression levels of MC4R mRNA existed tissue differences,and except for the liver,it existed breed differences,too.This research laid a basis for molecular marker assisted selection of growth traits,carcass traits,and meat quality traits of grazed goat.     

Isoenzymes of equine alkaline phosphatase

Alkaline phosphatase isoenzymes from small intestine, cecum, large colon, small colon, liver, kidney, leukocytes, and serum from ten clinically normal horses were defined by their sensitivities to L-phenylalanine, L-homoarginine, levamisole and heat, and by polyacrylamide gel disc electrophoresis. Readily identifiable isoenzymes occurred in small intestine, granulocytes, kidney, cecum, and large and small colon. By contrast, alkaline phosphatases from liver, lymphocytes, and serum could not be discriminated by this group of tests.     

陇东黑山羊肉品质及脂肪酸特征研究

为分析陇东黑山羊肉品质及脂肪酸特性,本研究在庆阳市随机选取3只12月龄陇东黑山羊公羊,测定其肉品质、营养成分和肌内脂肪酸。结果表明:背最长肌的pH在5.67～6.22,属于鲜肉的正常范围,剪切力为71.74 N,熟肉率为56.37%,肉色评分为3.33;背最长肌的水分含量(73.82%)高于前腿肌和后腿肌(P<0.05),背最长肌和前腿肌中的粗脂肪含量高于后腿肌(P<0.05),粗灰分和粗蛋白质含量在3个肌肉组织中差异均不显著;3个肌肉组织中单不饱和脂肪酸含量高于多不饱和脂肪酸(P<0.05),其中背最长肌中的油酸(C18:1n9c)含量为27.00%;多不饱和脂肪酸与饱和脂肪酸的比值(PUFA/SFA)在背最长肌中的含量为0.33。综合分析,胴体不同部位间营养成分和脂肪酸的组成及含量有一定差异性,背最长肌肉质最优,具有较好的食用价值和生产高档羊肉产品的潜力。     

CREB-H基因在猪不同组织中表达谱及肝脏中的表达规律

本研究旨在分析环腺苷酸应答元件结合蛋白H（cyclic AMP-responsive element-binding protein 3-like 3，CREB-H）在猪不同组织中的表达谱及其在马身猪和大白猪肝脏中的发育性表达规律。采用实时荧光定量PCR和Western blotting技术检测1日龄猪12个组织（心脏、肝脏、脾脏、肺脏、肾脏、胃、小肠、小脑、下丘脑、背最长肌、股肌和腰肌）中CREB-H基因的表达谱，以及CREB-H在1、30、60、90、120、150和180日龄马身猪和大白猪肝脏中的表达规律。结果显示，CREB-H基因mRNA在马身猪的12个组织中广泛表达，其中在肝脏和小肠中高表达；CREB-H蛋白在肝脏组织中的表达量显著高于其他组织（P<0.05），在心脏、脾脏和小脑中不表达。猪肝脏CREB-H基因mRNA和蛋白的发育表达受日龄、品种、品种与日龄相互作用的影响（P<0.01）。马身猪和大白猪肝脏中CREB-H基因mRNA和蛋白的表达量均在1日龄时达到最大值。在各发育阶段，马身猪CREB-H蛋白的表达量均极显著高于大白猪（P<0.01），且CREB-H主要在猪肝脏中表达。CREB-H在两猪种肝脏中的表达存在时空差异，可能与猪在不同发育期的脂质代谢能力有关，本试验结果为研究猪的脂质代谢调控机制提供一定的理论依据。     

贵州地方黄牛不同组织中PPARγ基因表达水平研究

试验旨在研究威宁牛、思南牛、关岭牛和黎平牛不同组织中过氧化物酶体增殖物激活受体γ（peroxisome proliferators-activated receptors γ,PPARγ）基因mRNA的表达差异。以这4个贵州地方品种黄牛为试验动物,提取心脏、肝脏、脾脏、肺脏、肾脏、脂肪和背最长肌组织的总RNA,设计PPARγ基因的实时荧光定量引物,以牛GAPDH基因为内参,应用实时荧光定量PCR技术检测PPARγ基因在4个品种不同组织中mRNA的相对表达量。结果显示,PPARγ基因在4个品种黄牛的心脏、肝脏、脾脏、肺脏、肾脏、脂肪和背最长肌组织中均有表达,但在脂肪组织中的表达量高于其他组织,且差异极显著（P<0.01）,在脾脏和肺脏中也有较高表达,而在肾脏、肝脏、心脏和背最长肌中表达量较低;不同品种PPARγ基因的表达在部分组织中存在差异。研究表明,PPARγ基因在不同组织中的表达量存在差异,可能与其在不同组织中的功能有关,而品种对于PPARγ基因的表达影响不大。