Different Months of MSTN Gene mRNA Expression Levels and their Correlation with Growth Traits in Five Xinjiang Local Sheep

To explore the different varieties of five Xinjiang local sheep myostatin (MSTN) expression levels and their correlation with growth traits (body weight,body height,body length,heart girth and cannon circumference).The MSTN mRNA expression level in different month(Birth,1 months,2 months,3 months,4 months,5 months) of muscle and fat in Altay sheep,Turpan Black sheep,Emil White sheep,Kazakh sheep,Bashbay sheep by quantitative Real-time PCR method,using SPSS 19.0 software to analyze the correlation with growth traits.The MSTN mRNA expression levels in muscle:The MSTN gene mRNA expression levels of Altay sheep,Kazakh sheep and Bashbay sheep at birth were lower than other months;The MSTN gene mRNA expression level of Emil White sheep in 3 months was significantly lower than other months (P<0.05);The MSTN gene mRNA expression levels of Altay sheep,Turpan Black sheep were the highest in 4 months;The MSTN gene mRNA expression level of Kazakh sheep was the lowest at birth,and was the highest in 5 months.The MSTN gene mRNA expression levels in fat:The MSTN gene mRNA expression level of Emil White sheep at birth was significantly lower than other months (P<0.05); The MSTN gene mRNA expression levels of Altay sheep,Emil White sheep and Bashbay sheep were the highest in 5 months,the MSTN gene mRNA expression level of Bashbay sheep in 5 months was significantly higher than other months (P<0.05);The MSTN gene mRNA expression level of Kazakh sheep was the highest in 2 month,it was significantly higher than 5 months (P<0.05).Correlation analysis revealed that the MSTN gene mRNA expression levels were mostly negatively correlated with growth traits.The MSTN gene mRNA expression levels in muscle and fat of Altay sheep were all negatively correlated with growth traits;The MSTN gene mRNA expression levels in muscle of Turpan Black sheep were negative correlated with body length and body height,others were positively;The MSTN gene mRNA expression levels in muscle of Emil White sheep were positive correlated with heart girth and cannon circumference;The MSTN gene mRNA expression levels in fat were positive correlated with body length,others were negatively correlation;The MSTN gene mRNA expression levels in muscle and fat of Kazakh sheep were positive correlated with body length,others were negatively correlation;The MSTN gene mRNA expression levels in muscle of Bashbay sheep were positive correlated with body length and cannon circumference,others were negatively correlation.The MSTN gene mRNA expression level was discrepancy in different month of different varieties,and was no fixed expression pattern;The MSTN gene mRNA expression levels were positively correlated with the part of the body size.The MSTN gene mRNA expression levels might have a direct relationship with the growth traits and bone growth.     

不同月龄红河黄牛抗病基因组织mRNA表达量分析

为了解天然抗性相关巨噬细胞蛋白1基因(SLC11A1)、Toll样受体2(TLR2)、Toll样受体4(TLR4)、γ-干扰素基因(BoIFN-γ)、α-干扰素基因(BoIFN-α)和主要组织相容性复合物Ⅱ类基因DRB3(BoLA-DRB3)6种抗病相关基因,在红河黄牛不同组织的表达分布情况,根据GenBank收录的基...     

新西兰白兔MSTN基因克隆、生物信息学分析及表达谱构建

肌肉生长抑制素（MSTN）是骨骼肌发育的主要调控因子,为获得新西兰白兔MSTN基因序列及其表达规律,试验通过RT-PCR技术从新西兰白兔腿肌组织中扩增并克隆得到MSTN基因序列,利用在线网站对其进行生物信息学分析,并采用实时荧光定量PCR技术检测MSTN基因在新西兰白兔同一时期不同组织及不同时期腿肌组织中的表达量。结果显示,新西兰白兔MSTN基因CDS区序列长1 128 bp,编码375个氨基酸,其核苷酸序列与GenBank上公布的人、猪、马、小鼠、犬、牛和鸡的核苷酸序列相似性分别为95.9%、94.9%、94.9%、91.7%、91.5%、91.3%及84.2%。新西兰白兔MSTN蛋白属于酸性、亲水性分泌蛋白,且具有不稳定性,不含跨膜结构,含有1个信号肽,主要分布在内质网和线粒体中。MSTN蛋白二级结构和三级结构预测结果一致,以无规则卷曲为主,其次是α-螺旋、延伸链,为混合型蛋白。蛋白质互作关系预测发现,MSTN蛋白与PAX7、MYOG、IGF1、FST、Smad3及Smad2等与肌肉生长发育有关的蛋白之间存在相互作用。不同组织表达分析结果表明,新西兰白兔MSTN基因在心脏、肝脏、脾脏、肺脏、肾脏、腿肌、子宫和胃中均有表达,在腿肌中表达量最高,极显著高于其他组织（P<0.01）,其次是肾脏,在肝脏中的表达量最低。不同时期腿肌组织中的表达量分析结果显示,新西兰白兔MSTN基因在180日龄腿肌组织中的表达量显著高于90和240日龄（P<0.05）。研究结果为深入探讨MSTN基因对家兔肌肉生长发育的调控机制奠定了基础。     

海南黑山羊MSTN基因的表达差异与发育性变化研究

采用SYBR Green实时荧光定量PCR技术检测了MSTN基因在2、3、4月龄和12月龄海南黑山羊腿肌、腰肌和背肌组织中mRNA表达发育性变换规律,为揭示MsTN基因的生物学功能及其对肉质的表达调控机理提供参考依据。结果表明,在海南黑山羊的四个生长阶段,MSTN基因均在背肌里的表达量最高,腿肌里的表达量较低,表达模式为：背肌〉腰肌〉腿肌;腿肌和腰肌在2月龄时,表达量较低,3月龄时极显著升高（P〈0．01）,4月龄时又极显著下降（P〈0．01）,12月龄时又极显著升高（P〈0．01）;表达模式为：12月龄〉3月龄〉4月龄〉2月龄;背肌在2月龄时表达量也较低,3月龄时极显著升高（P〈0．01）,达到最高水平,4月龄时又极显著下降（P〈0．01）,12月龄时有所上升,但差异不显著（P〉0．05）,表达模式为：3月龄〉12月龄〉4月龄〉2月龄。     

东佛里生羊MSTN基因多态性及其与生长性状的关联分析

试验旨在研究肌肉生长抑制素(myostatin,MSTN)基因多态性及其对东佛里生羊生长性状的影响。从324只东佛里生羊全血中提取DNA,对MSTN基因外显子序列和非翻译区(untranslated region,UTR)序列进行PCR扩增,采用一代测序技术鉴定单核苷酸多态性(single nucleotide polymorphism,SNP)位点,分析不同位点的遗传多态性,并将不同基因型与东佛里生羊的生长性状进行关联分析。结果显示,在东佛里生羊MSTN基因外显子中未检测到突变;在3'-UTR区的272 bp处存在G→A单碱基突变,该位点只检测到AG和AA基因型,且AA基因型频率高于AG基因型;在5'-UTR区的176 bp处存在C→A突变,该位点只检测到CC和AC基因型,且CC基因型频率高于AC基因型。遗传多样性参数分析结果表明,东佛里生羊群体MSTN基因3'-UTR-272和5'-UTR-176位点的纯合度分别为0.827和0.887,杂合度分别为0.173和0.113,2个位点的多态性均较低(PIC<0.25),其群体基因型分布均符合哈代-温伯格平衡(P>0.05)。关联分析发现,3'-UTR-272位点AG基因型体斜长显著高于AA基因型(P<0.05),5'-UTR-176位点CC基因型胸围显著高于AC基因型(P<0.05),其余指标均无显著差异(P>0.05)。综上,东佛里生羊MSTN基因3'-UTR和5'-UTR突变位点可作为其目标性状选育的分子标记。     

MSTN基因在1~6月龄哈萨克羔羊肌肉组织中表达量及其与生长指标相关性分析

试验旨在分析MSTN基因在1~6月龄哈萨克羔羊肌肉组织中表达量及其与各项生长指标的相关性,为哈萨克羊的良种选育提供科学依据.随机选取5只哈萨克公羔羊,每月定时在羔羊活体腿部取肌肉组织,软尺测量体长、体高、体重、管围、胸围等各项生长指标.实时荧光定量 PCR分析1~6月龄哈萨克羔羊肌肉组织中MSTN的表达量,相关性分析MSTN基因在羔羊肌肉中的表达量与各项生长指标的关联性.结果显示:哈萨克羔羊各项生长指标在1、2月龄快速增长,体重从4月龄开始增重缓慢,体长、体高、胸围在3月龄出现增长缓慢,管围在3月龄起基本停止增长.MSTN基因在1~4月龄哈萨克羔羊肌肉组织中表达量均无显著性差异(P>0.05),呈现略微上升的趋势,5月龄达到最高峰,6月龄迅速下降.MSTN 基因在肌肉中表达量与体长呈显著负相关 (P<0.05),相关系数为-0.472,与体重、胸围呈弱负相关,与管围、体高呈弱正相关.     

实时荧光定量PCR检测多重耐药大肠杆菌AcrA基因mRNA表达水平

体外单一药物诱导标准大肠杆菌ATCC25922,获得了耐氯霉素（CHL,MIC≥256 mg/L）、耐环丙沙星（CIP,MIC≥256 mg/L）、耐水杨酸钠（SAL,MIC≥275 mg/L）、耐四环素（T,MIC≥512 mg/L）的菌株。选取ATCC25922株,4株中度耐药菌SAL（175）、CHL（64）、T（64）、CIP（128）,4株高度耐药菌SAL（275）、CHL（256）、T（512）、CIP（256）,和临床分离的3株耐药大肠杆菌H1、H9、H10,通过实时荧光定量RT-PCR方法检测外输泵AcrA基因的mRNA表达水平。系统自动分析软件显示,Ct值与标准质粒浓度的对数之间存在良好的线性关系,回归系数为0.992。检测结果表明：H10、CHL（256）、CIP（256）拷贝数最高,达1015拷贝/μL;T（512）、H9、T（128）、SAL（275）、H1为1014拷贝/μL;CHL（128）、T（64）、CHL（64）为1013拷贝/μL;ATCC25922为1012拷贝/μL。不同程度耐药株AcrA基因的表达量不同,临床分离株和诱导的各高耐药菌株均比ATCC25922株表达量高,且差异极显著（P≤0.01）。这说明不同耐药程度的11株菌cDNA的量存在差异,AcrA基因转录水平与耐药水平成正相关。     

绵羊CLPG和MSTN基因多态性及其与生长性状的关联分析

试验旨在探究CLPG(Callipyge)与MSTN(Myostatin)基因作为绵羊生长性状候选基因的可能性,调查与绵羊生长性状相关的分子遗传标记。本试验以133只澳洲白羊×杜泊羊×湖羊杂交绵羊为研究对象,利用PCR产物直接测序及PCR-RFLP技术检测CLPG与MSTN基因的单核苷酸多态性,然后通过SPSS22.0软件GLM统计模型分析多态位点不同基因型及聚合基因型与绵羊生长性状的关联性。测序结果表明,CLPG基因STS序列232bp处检测到C→T突变位点C1,MSTN基因3'UTR区检测到G→A突变位点M1。PCR-RFLP分析显示,C1位点表现为2种基因型:CC和CT;M1位点表现为2种基因型:GG和GA。关联分析表明,C1位点与绵羊背膘厚和眼肌面积显著或极显著相关(P<0.05;P<0.01),M1位点与绵羊体重、管围、背膘厚和眼肌面积显著或极显著相关(P<0.05;P<0.01)。聚合基因型对体重、背膘厚和眼肌面积影响极显著(P<0.01)。研究揭示CLPG与MSTN基因多态性及其聚合基因型对绵羊生长性状具有显著影响,C1和M1位点可以考虑作为绵羊生长性状的有效遗传标记。     

吐鲁番黑羊MSTN/Smad信号通路基因表达的发育性变化

为探讨MSTN/Smad信号通路基因对吐鲁番黑羊肌肉生长发育的影响,采用实时定量PCR法,分别对1～6月龄吐鲁番黑羊的腿肌和尾脂MSTN/Smad信号通路基因进行检测.结果表明：MSTN/Smad信号通路基因在腿肌和尾脂组织中均有表达,MSTN/Smad信号通路基因在吐鲁番黑羊不同生长阶段的腿肌和尾脂中的表达没有出现随月龄的增加而一直增加或下降的趋势.     

鸡、鹌鹑及其杂交禽胚胎肌肉发育过程中MSTN和p21基因的表达规律

旨在研究MSTN和p21基因在鸡、鹌鹑和杂交禽胚胎肌肉发育过程中的表达规律,比较这2个基因在杂交禽与亲本鸡和鹌鹑中的差异表达,探讨MSTN和p21基因与肌肉发育的关系。通过人工受精获得鸡(♂)与鹌鹑(♀)杂交禽蛋,与鸡蛋、鹌鹑蛋按照鸡标准孵化条件同批入孵,采集第7～17天活胚胸肌组织,应用实时荧光定量PCR技术检测MSTN和p21基因在3个物种中每一天mRNA的相对表达量。MSTN和p21基因在胚胎肌肉发育的第7～17天均有表达,这2个基因mRNA的表达在鸡胚中第9天到达第1次峰值,在鹌鹑中第7天到达第1次峰值,后都随胚胎的发育表达水平上升,并维持相对稳定的高水平表达。杂交禽的表达规律与鸡一致,也在第9天达到峰值,而p21mRNA表达极显著高于鸡和鹌鹑(P<0.01)。MSTN mRNA表达规律与成肌细胞退出细胞周期的时间规律一致;在胚胎肌肉发育过程中,MSTN基因特异性上调p21的表达。     

新疆也木勒白羊肌肉生长抑制素基因在不同生长时期的表达研究

试验采用实时荧光定量PCR技术测定也木勒白羊肌肉生长抑制素(myostatin,MSTN)基因在不同生长时期(0～6月龄)的表达量,采用2^－ΔΔCt分析不同生长阶段表达量的差异,旨在掌握也木勒白羊不同生长时期MSTN基因的表达规律,为也木勒白羊肌肉生长性状的选育提供遗传学资料.结果发现,MSTN基因在3月龄也木勒白羊肌肉中的表达量显著低于其他月龄(P<0.05),其余各月龄差异不显著(P>0.05);MSTN在脂肪中的表达量0月龄相对较低,随月龄增长,表达量升高,在5月龄达到最高,后随月龄增长表达量降低.MSTN基因在肌肉和脂肪中的表达量随着月龄增长有一定的规律可循,其表达规律可为也木勒白羊肉用性能的选育提供一定的科学依据.     

新疆巴什拜羊肌肉生长抑制素基因DNA甲基化与mRNA表达量分析

为了探究肌肉生长抑制素（myostatin,MSTN）基因在肌肉和脂肪中的DNA甲基化模式及mRNA表达水平,试验以5月龄巴什拜羊羔羊为研究对象,采用亚硫酸氢盐测序法（bisulfite sequencing PCR,BSP）检测MSTN基因启动子区和第1外显子甲基化模式,并通过实时荧光定量PCR检测MSTN基因在巴什拜羊股二头肌、股三头肌、半腱肌、半膜肌、背最长肌和尾脂中的mRNA表达水平。结果显示,肌肉组织甲基化概率高于脂肪组织,其中,股二头肌、股三头肌、半腱肌、半膜肌、背最长肌和尾脂的甲基化概率分别为74.2%、74.2%、83.2%、83.7%、82.1%和25.3%,MSTN基因在股二头肌、股三头肌、半腱肌、半膜肌、背最长肌中的表达水平显著低于尾脂（P < 0.05）,股二头肌、股三头肌、半腱肌、半膜肌和背最长肌之间差异不显著（P > 0.05）,巴什拜羊肌肉和脂肪MSTN基因DNA甲基化水平与MSTN基因表达量呈显著负相关（r=－0.886,P < 0.05）。     

Analysis of DNA Methylation and mRNA Expression Level of MSTN Gene in Xinjiang Bashiby Sheep

In order to investigate DNA methylation and expression levels of myostatin (MSTN) gene in mscule and fat, 5 months of age of Bashiby sheep were selected, the promoter region and exon 1 methylation levels of MSTN gene was analyzed using bisulfite sequencing PCR (BSP). Real-time PCR was used to detect the expression level of MSTN gene in biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle and tail fat of Bashiby sheep. The results showed that the methylation probability of muscle was higher than fat, the methylation probability of biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle and tail fat were 74.2%, 74.2%, 83.2%, 83.7%, 82.1% and 25.3%, respectively. The expression levels of MSTN gene in biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle were significantly lower than tail fat in Bashiby sheep (P < 0.05), but there were no significant difference among biceps femoris, femoral triceps, semitendinosus, semimembranosus and longissimus dorsi muscle (P > 0.05).The DNA methylation was negatively correlated with the expression levels in muscle and fat of Bashiby sheep (r=-0.886, P < 0.05).     

Polymorphism of CLPG and MSTN Genes and their Association Analysis with Growth Traits in Sheep

The study was conducted to explore the possibility that CLPG (Callipyge) and MSTN (Myostatin) genes which could be the candidate genes of sheep growth traits, and investigate the molecular genetic markers related to sheep growth traits.133 (Austrilian White sheep×Dorper sheep×Hu sheep) hybid-sheep were chosen as subjects, the technology of direct sequencing of PCR products and PCR-RFLP were used to detect the single nucleotide polymorphism of CLPG and MSTN genes, then the association of the SNPs different genotypes and combined genotypes with sheep growth traits were analyzed by the GLM statistical model of SPSS 22.0.Sequencing results showed that the SNP of C/T which called C1 was detected at position 232 bp of the STS sequence in CLPG gene.The SNP of G/A which called M1 was detected in the 3'UTR of MSTN gene.PCR-RFLP analysis showed that two genotypes CC and CT were in C1 site, two genotypes GG and GA were in M1 site.Association analysis revealed that C1 site was significantly or extremely significantly associated with backfat thickness and loin muscle area (P<0.05;P<0.01), M1 site was significantly or extremely significantly associated with body weight, tube girth, backfat thickness and loin muscle area (P<0.05;P<0.01).Meanwhile, the combined genotype was extremely significantly associated with body weight, backfat thickness and loin muscle area (P<0.01).The conclusions indicated that SNPs and combined genotype of CLPG and MSTN genes had effects on growth traits in sheep.C1 and M1 sites could be considered as effective genetic markers for sheep growth traits.     

Expression of MSTN and MyoG Genes in Zi and Rhine Goose and their Correlation with Carcass Traits

The aim of this study was to investigate the effects of MSTN and MyoG genes on goose skeletal muscle growth.In this study,MSTN and MyoG genes expression were detected in breast and leg muscle of Zi and Rhine goose by Real-time PCR,and the correlations between genes expression levels and carcass traits were investigated.The results showed that the breast muscle weight and breast muscle rate of Rhine goose were extremely significant higher than Zi goose (P<0.01).MSTN and MyoG mRNA expression in breast muscle of Zi goose were significantly higher than that of Rhine goose,and the mRNA level of MSTN in leg muscle of Rhine was extremely significant higher than that of Zi goose (P<0.01),there was no significant difference of MyoG mRNA between Zi goose and Rhine goose (P>0.05).There was extremely significant difference between MSTN mRNA expression in breast muscle and leg muscle of Zi goose (P<0.01).MSTN mRNA expression in leg muscle was significantly higher than that of breast muscle of Rhine goose (P<0.05).There was extremely significant difference between MyoG mRNA expression in breast muscle and leg muscle of Zi goose and Rhine goose (P<0.01).There was a extremely significant negative correlation between MSTN mRNA expression in breast muscle with body weight,breast muscle weight and breast muscle percentage (P<0.01).There was a extremely significant positive correlation between MyoG mRNA expression in breast muscle and leg muscle with body weight (P<0.01).MSTN and MyoG gene might have positive and negative regulation effect on muscle growth.     

籽鹅和莱茵鹅MSTN和MyoG基因表达及其与屠宰性状的相关分析

本研究旨在探讨肌肉生成抑制素(myostatin,MSTN)和肌细胞生成素(myogenin,MyoG)基因对鹅骨骼肌生长发育的影响。试验以莱茵鹅和籽鹅为研究对象,采用实时荧光定量PCR方法测定MSTN、MyoG基因在籽鹅和莱茵鹅胸肌、腿肌中的差异表达情况,运用统计软件对基因表达情况与屠宰性状间的相关性进行分析。结果显示,莱茵鹅胸肌重和胸肌率极显著高于籽鹅(P<0.01),籽鹅胸肌MSTN、MyoG mRNA表达量极显著高于莱茵鹅(P<0.01);在腿肌中,籽鹅MSTN mRNA表达量极显著低于莱茵鹅(P<0.01),籽鹅与莱茵鹅MyoG mRNA表达量间无显著差异(P>0.05)。同一品种MSTN mRNA表达水平也存在差异,在籽鹅胸肌和腿肌中表达量存在极显著差异(P<0.01);在莱茵鹅胸肌和腿肌中表达量存在显著差异(P<0.05)。就MyoG mRNA而言,在籽鹅和莱茵鹅胸肌和腿肌中表达量存在极显著差异(P<0.01)。MSTN基因表达与屠宰性能相关性分析表明,胸肌中MSTN mRNA表达量与活重、胸肌重和胸肌率呈极显著负相关(P<0.01);胸肌和腿肌中MyoG mRNA表达量与活重呈极显著正相关(P<0.01)。说明MSTN和MyoG基因可能对肌肉生长分别起正负调控作用。