鸡apoA5基因单核苷酸多态性及其与屠体性状的关联研究

以丝羽乌骨鸡和隐性白洛克正反交产生的F2代为实验群体, 采用PCR-SSCP和DNA测序的方法检测鸡载脂蛋白A5(apoA5)基因的单核苷酸多态性(SNPs), 并将所发现的SNPs与体重、胸肌重、腿肌重、心脏重、肝脏重和腹脂重等屠体性状进行关联分析。结果发现, 鸡apoA5基因5′-调控区C-169T, 外显子2 C600T、T635C, 外显子3 C841G、C914T、C1142G、C1394T共7个突变位点。其中外显子2突变位点C600T、T635C对12周龄腹脂重、腹脂率、肝脏重和心脏重有显著影响(P<0.05), 根据PCR-SSCP的结果将其分为6种基因型(AA, AB, AC, BB, BC, CC): 其中CC型个体的腹脂重和腹脂率显著高于AA型、AB型、AC型、BB型、BC型 (P<0.05); AC型个体的肝脏重显著低于AA型、AB型、BB型、BC型和CC型的肝脏重(P<0.05); BC型个体的心脏重显著低于BB型的心脏重(P<0.05)。     

鸡Myostatin基因单核苷酸多态性的群体遗传学分析

肌肉生长抑制素是控制骨骼肌生长发育的重要细胞因子,采用PCR－SSCP和测序的方法发现了5个位于Myostatin基因5′－和3′－调控区的单核苷酸多态性位点,对北京油鸡、白耳鸡、石歧杂、矮小黄鸡、小型黄鸡、惠阳胡须鸡、隐性白羽鸡、海兰、AA鸡等不同鸡种的该单核苷酸多态性分析结果表明：Myostatin基因的5′调控区引物P60／P61扩增片段多态性是由3个核苷酸的改变而产生的[分别是G→A（304位）、A→G（322位）、G→（344位）],引物P93／P94扩增片段的多态性是由G→A（167位）突变造成的,引物P117。P118PC扩增片段多态性是由T→C（177位）造成的。3′调控我引物P80／P81扩增片段多态性是由第7263位A突变为T造成的,引物P76／P77扩增片段多态性是由A→G（6935位）造成的。不同鸡种群体遗传学分析表明,5′－调控区引物60／P61扩增片段多态性片段多态性是由A→G（6935位）造成的。不同鸡种群体遗传学分析表明,5′－调控区引物P60／P61扩增片段多态性位点在北京油鸡的基因型频率分布与其他的品种有很大的差异,其BB型频率为0．700,AA基因型频率仅为0．033,而其他鸡种中以A基因优势;对于引物P93／P94,品种间的基因型频率差异极显著（P＜0．01）,北京油鸡和AA鸡的EE型频率鸡种中以A基因占优势;对于引物P93／P94,品种间的基因型频率差异极显著（P＜0．01）,北京油鸡和AA鸡的EE型频率低于其他品种,白耳鸡和海兰蛋鸡以EE型为主,其频率高于其他品种;3′－调控区引物P80／P81多态怀位点在9个鸡种中都是等位基因C占优势。引物P76／P77,总体上MM型的频率较低,杂合子MN型的频率较高。     

鸡lmbr1基因单核苷酸多态与生长及屠体性状的关联性

Lmbr1基因是脊椎动物肢体发育的关键候选基因, 该基因对畜禽生长及屠体性状的影响尚未见报道. 本研究以lmbr1基因作为控制鸡生长及屠体性状的候选基因, 以丝羽乌骨鸡和白洛克肉鸡杂交产生的F2资源鸡群为实验群体. 采用单链构象多态性(SSCP)结合测序方法在外显子16检测到T/C和G/A突变各一, 在内含子5检测到一个A/C突变. 在亲代检测到突变位点后, 进一步进行F2代鸡群的基因型检测. 方差分析结果显示鸡lmbr1基因外显子16的T/C多态与全净膛率、肌胃率、胫爪率和胫围显著相关, 内含子5的A/C多态与胫爪率、肝脏比率和头颈重显著相关, 但两个位点都与其他生长及屠体性状的相关不显著. 研究结果表明, 鸡lmbr1基因应是控制这些生长及屠体性状的主效基因或与控制这些性状的主效基因紧密连锁.     

鸡MC4R基因的SNPs及其与屠体性状的相关研究

黑素皮质素受体(MC4R, melanocortin-4 receptor)基因的突变与猪、鼠和人等的食欲、肥胖、生长等性状有关, 而鸡MC4R基因的功能却知之甚少. 利用PCR-SSCP(single strand conformation polymorphism)和DNA测序的方法, 对资源家系F₂代鸡群MC4R基因多态性进行了分析, 发现存在4个单核苷酸多态(SNPs, single nucleotide polymorphisms)位点. 其中, 在MC4R基因5′调控区-524 nt发生了碱基的转换突变(C→T), 导致突变型基因比野生型基因多了一个NF-E2和一个cap转录因子结合位点; 在MC4R编码区(61 nt)发生了碱基的错义突变(G→A), 导致此处蛋白质的氨基酸由甘氨酸变为精氨酸; 在MC4R编码区315和336 nt发生了碱基的颠换突变(G→T)和转换突变(C→T), 这两个突变为同义突变. 通过最小二乘分析SNPs与屠体性状的关系, 结果是突变的BB, DD和FF等基因型与鸡的体重、全净膛重(或半净膛重)、腿肉重等存在显著(P<0.05)或极显著(P<0.01)的关系, 但与腹脂重不显著. 结果表明, MC4R基因可以作为影响和控制鸡体重、生长等屠体性状的主要候选基因.     

兔黑素皮质素受体4(MC4R)基因多态性及其与体重及屠体性状的关联研究

以哈尔滨白兔、天府黑兔、比利时兔、齐卡巨型兔以及加利福尼亚兔5个肉兔品种共220个个体为研究对象, 采用PCR-SSCP技术和克隆测序技术进行SNPs检测和基因型的分析。结果发现兔MC4R基因237位发生转换突变(A→G), 5个品种内的AA基因型和A等位基因频率均分别高于AG基因型和G等位基因, 而哈尔滨白兔、比利时兔以及齐卡巨型兔3个品种兔的AG基因型频率和G等位基因频率明显高于天府黑兔和加利福尼亚兔。单位点基因型与屠体性状、饲料转化率以及熟肉率的最小二乘分析表明; AG基因型与兔的体重、全净膛重以及饲料转化率有显著的关系(P<0.05), 而与熟肉率的关系不显著(P>0.05)。结果表明, MC4R基因可以作为影响和控制兔体重及屠体性状的候选基因。     

鸭前胰岛素原基因多态性及其与屠体性状的关联分析

以384只北京鸭 (Z2系、Z4系、Z2×Z4杂交系)和樱桃谷鸭为材料, 利用PCR-SSCP结合测序技术, 对前胰岛素原基因外显子2与部分内含子的多态性进行了研究, 并分析对屠体性状的遗传效应。结果发现存在2个单核苷酸突变位点, 即在第179位和第195位分别发生了T→C和C→T的突变。适合性χ2检验结果表明, 北京鸭各品系和樱桃谷鸭均处于Hardy-Weinberg平衡状态(P>0.05)。最小二乘分析SNPs与屠体性状的关系表明, 在北京鸭3个品系中, 基因型 BB 在胴体重、全净膛重和胸肌重上极显著(P<0.01)高于基因型AA和AB, 在腿肌重和皮脂重上极显著(P<0.01)高于基因型AB; 基因型AA在皮脂率和全净膛重上极显著(P<0.01)和显著(P<0.05)高于基因型AB。而对于樱桃谷鸭, 只有AB型在皮脂重和腹脂重上显著(P<0.05)高于基因型AA。研究结果表明, 鸭前胰岛素原基因多态性与鸭的部分屠体性状存在显著相关性, 且B等位基因有利于增加鸭的胴体重和胸肌重。     

绵羊Myostatin基因敲除载体的构建

根据Myostatin基因突变可导致肌肉量激增而产生"双肌"表型的特点,构建绵羊Myostatin基因置换型敲除栽体.利用LA-PCR技术成功地扩增得到绵羊Myostatin基因同源臂序列,其中同源长臂4.9kb,包括全部的exonl,intronl,exon2及部分启动子和大部分intron2;同源短臂1.1kb,包括部分exon3和31非翻译区序列,将二者连入PloxpⅡ正负筛选敲除骨架载体,利用骨架载体上Neo基因替代Myostatin基因的exon3,从而成功构建专门针对Myostatin第3外显子区域缺失的置换型敲除载体PloxpⅡ-OVIS-MSTN.酶切和测序鉴定证明载体构建正确,为后续获得绵羊Myostatin基因缺失型体细胞株奠定试验基础.     

猪UCP5基因克隆表达特性及与屠体性状的关联分析

通过3′和5′cDNA末端快速扩增(RACE)获得了猪UCP5基因全长cDNA序列,通过不同品种猪UCP5基因编码区和调控区单核苷酸多态分析,发现 -1 567 bp处存在A→T的突变,从而导致转录因子CdxA、HNF-1、Sox-5、GATA-2结合位点发生了改变．在金华×皮特兰F2代资源群中,使用PCR限制性片段长度多态性(RFLP)方法,对524头个体进行基因型测定,并与屠体性状关联分析,结果发现,AA型活体重极显著高于BB型(P < 0.01),AB型活体重显著高于BB型(0.01 < P < 0.05),AA型的后腿肌肉重极显著高于BB型(P < 0.01),AA型的后腿脂肪重显著高于BB型(0.01 < P < 0.05)．同时,通过实时荧光定量PCR获得了UCP5基因在初生仔猪心、肝、脾、肺、肾、眼肌、腹脂和大脑等不同组织中基因表达的特性,结果表明,UCP5在猪的各种组织均有表达,在脑和肺中表达量最多．t检验发现,初生达兰猪眼肌中UCP5的表达量显著高于金华猪．     

鸡类胰岛素生长因子-Ⅱ基因单核苷酸多态与生长、屠体性状相关性的研究

以鸡类胰岛素生长因子-Ⅱ（IGF-Ⅱ）基因作为控制鸡生长、屠体性状主基因的侯选基因,以明显肉鸡和丝毛乌骨鸡杂交产生的F2代鸡群为实验材料。通过PCR-RFLP方法对IGF-Ⅱ基因进行多态性检测。在该基因外显子2中发现一处碱基突变,可由内切酶Aci-I酶切证实,但未导致氨基酸改变。对F2鸡群个体进行了基因型鉴定,结果经方差分析显示;该基因对一些生长、屠体性状有显著性影响（如胸角宽、腺胃重、半净膛重等）。表明该基因可能调控鸡生长、屠体性状的表达,或与控制生长、屠体性状的主要因连锁。     

鸡Myf6基因遗传多态性及其遗传效应研究

采用测序和单链构象多态(SSCP)的方法分析了Myf6基因在5个优质肉鸡纯系和3个杂交配套系中的遗传分布、遗传变异及群体杂合性等群体遗传信息, 并分析了Myf6基因对胴体性状及肉质性状的遗传效应。结果发现该基因的CDS区域非常保守, 仅在外显子1的47位处发生点突变由G→A, 由于人工选育的原因, 该基因不同基因型的分布在2个优质肉鸡纯系中偏离了Hardy-Weinberg平衡, 并且杂合度较低, 遗传一致性较高, 但在3个杂交配套系中仍都服从Hardy-Weinberg平衡, 杂合度较高, 达到了配套杂交的目的。野生型A基因可显著地提高活重、屠体重、胸肌重和腿肌重(P <0.05), 同时也会降低肉的嫩度。具体地讲, A基因对增加活重、屠体重、胸肌重和腿肌重的加性效应值分别为45.54 g, 41.03 g, 6 g和9.775 g; 对增加肌纤维直径和肌纤维密度的加性效应值分别为1.025 mm和-29.99 mm2。     

Single nucleotide polymorphisms in chicken lmbr1 gene were associated with chicken growth and carcass traits

Lmbr1 is the key candidate gene controlling vertebrate limb development, but its effects on animal growth and carcass traits have never been reported. In this experiment, lmbr1 was taken as the candi-date gene affecting chicken growth and carcass traits. T/C and G/A mutations located in exon 16 and one A/C mutation located in intron 5 of chicken lmbr1 were detected from Silky, White Plymouth Rock broilers and their F2 crossing chickens by PCR-SSCP and sequencing methods. The analysis of vari-ance (ANOVA) results suggests that T/C polymorphism of exon 16 had significant association with eviscerated yield rate (EYR), gizzard rate (GR), shank and claw rate (SCR) and shank girth (SG); A/C polymorphism of intron 5 was significantly associated with SCR, liver rate and head-neck weight (HNW), while both sites had no significant association with other growth and carcass traits. These results demonstrate that lmbr1 gene could be a genetic locus or linked to a major gene significantly affecting these growth and carcass traits in chicken.     

A-FABP基因多态性与肉鸡生长和体组成性状的关联

为探讨A-FABP基因多态性对肉鸡生长和体组成性状的影响,文章选用肉鸡高、低脂双向选择品系第十世代肉仔鸡为实验材料,采用测序、PCR-RFLP、PCR-LP、DHPLC方法进行基因多态性检测和个体基因型分析,通过对8个SNPs进行连锁不平衡分析并选择5个标签SNPs构建单倍型,进而利用单位点和单倍型分别与鸡生长和体组成性状进行关联分析。结果表明,7个SNPs(除SNP 5以外)及单倍型同时对鸡肌胃重、肌胃率有显著影响(P<0.05),而对生长和其他体组成性状无显著影响(P>0.05)。鉴于不同物种A-FABP基因现有的功能研究均没有发现其对消化系统有重要作用,因此该基因是否为影响肉鸡肌胃重和肌胃率的主效基因还有待于进一步研究。     

Isolation and imprinting analysis of the porcine DLX5 gene and its association with carcass traits

Imprinted genes play important roles in mammalian growth and development. However, reports on imprinted genes are limited in livestock. In this study, the complete ORF containing 289 amino acids of the porcine DLX5 gene was obtained. A C-to-T SNP mutation in exon 1 of the DLX5 gene was used to detect imprinting status with an RT-PCR/RFLP test (using HhaI) in eight heterozygous pigs from a population of Large White × Meishan F₁ hybrids. Imprinting analysis showed that the porcine DLX5 gene was maternally expressed in skeletal muscle, fat, lung, spleen, stomach and small intestine, but not imprinted in heart, liver, kidney, uterus, ovary, testicle or pituitary. A PCR–RFLP test was also used to detect the polymorphism in 310 pigs of a Large White × Meishan F₂ resource population. The statistical results showed significant association ( P  < 0.01) of the genotypes and fat meat percentage, carcass length, bone percentage, 6–7 rib fat thickness, average backfat thickness, thorax-waist fat thickness and buttock fat thickness.     

Single nucleotide polymorphisms in chicken lmbr1 gene were associated with chicken growth and carcass traits

Lmbr1 is the key candidate gene controlling vertebrate limb development, but its effects on animal growth and carcass traits have never been reported. In this experiment, lmbr1 was taken as the candidate gene affecting chicken growth and carcass traits. T/C and G/A mutations located in exon 16 and one A/C mutation located in intron 5 of chicken lmbr1 were detected from Silky, White Plymouth Rock broilers and their F₂ crossing chickens by PCR-SSCP and sequencing methods. The analysis of variance (ANOVA) results suggests that T/C polymorphism of exon 16 had significant association with eviscerated yield rate (EYR), gizzard rate (GR), shank and claw rate (SCR) and shank girth (SG); A/C polymorphism of intron 5 was significantly associated with SCR, liver rate and head-neck weight (HNW), while both sites had no significant association with other growth and carcass traits. These results demonstrate that lmbr1 gene could be a genetic locus or linked to a major gene significantly affecting these growth and carcass traits in chicken. Supported by the State Major Basic Research Development Program (Grant No. G20000161) and Beijing Natural Science Foundation (Grant No. 5011002)     

Lack of association of bovine MHC class I alleles with carcass and reproductive traits

The present study was carried out to examine whether a relationship between bovine major histocompatibility complex (BoLA) class I alleles and carcass traits or reproductive performance exists in Braunvieh and Fleckvieh A1 (artificial insemination) bulls. The influence of BoLA class I (BOLA-A) alleles on deregressed breeding values for net growth rate, carcass index and thigh volume was assessed in Braunvieh crosses and Fleckvieh bulls with a gene substitution model. The reproductive traits: non-return rate and interval between first and last insemination of daughters (female fertility), as well as non-return rate of inseminated cows (male fertility), were only investigated in Fleckvieh animals. No influence of the BOLA-A region on the traits evaluated could be demonstrated. An improper, i.e. less restrictive analysis would have led to spurious results.     

Polymorphisms in the ovine myostatin gene (MSTN) and their association with growth and carcass traits in New Zealand Romney sheep

Myostatin is a regulator of myogenesis and has been implicated in the regulation of adiposity and in controlling the structure and function of tendons. Polymerase Chain Reaction Single-Stranded Conformational Polymorphism (PCR-SSCP) analysis of intron-1 was used to identify five variants (designated A-E) of the myostatin gene ( MSTN ). The effect of this genetic variation on growth and carcass traits was investigated in 517 Romney male lambs from 17 sire-lines, born on a South Island New Zealand farm. General linear mixed effect models revealed that the presence of allele A in a lamb's genotype was associated with decreased leg, loin and total yield of lean meat, whereas the presence of allele B was associated with increased loin yield and proportion loin yield (loin yield divided by total yield expressed as percentage). The effect of the number of allele copies present was investigated, and it was found that the absence of A , or the presence of two copies of B , was associated with increased mean leg yield, loin yield and total yield. Two copies of B were also associated with a decrease in proportion of shoulder yield, whereas two copies of A were associated with a decrease in proportion of loin yield. Associations with allele C were not detected. No associations of MSTN variation with birth weight, weaning weight, pre-weaning growth rate, draft age and hot carcass weight (H-W) were detected. These results suggest that variation in ovine MSTN is associated with meat production, but not birth weight or growth rate in New Zealand Romney sheep.