奶牛性别鉴定的研究与应用

 以控制部分绵、山羊品种高繁殖力的BMPR IB、BMP15和GDF9基因为候选基因,采用PCR RFLP方法分析闽东山羊BMPR IB,BMP15 和GDF9 基因多态性与繁殖性状的关系。研究发现：多胎品种闽东山羊及南江黄羊在BMPR IB 基因的相应位置上并未发生与Booroola Merino 羊相同的突变,同时也未检测到BMP15 的FecXI, FecXH , FecXB基因及GDF9的FecGH 基因,因此排除了这5个突变位点影响闽东山羊高繁殖力性状的可能性。然而,由于闽东山羊的BMPR IB、BMP15、GDF9 的全基因序列信息的缺乏,尚不能完全断定3个基因对闽东山羊高繁殖力性状没有影响。     

BMPR-IB、BMP15、GDF9基因作为福清山羊高繁殖力候选基因的研究

以控制部分绵、山羊品种高繁殖力的BMPR-IB、BMP15和GDF9基因为候选基因,采用PCR-RFLP方法分析福清山羊BMPR-IB、BMP15和GDF9基因多态性与繁殖性状的关系。结果表明:多胎品种福清山羊及南江黄羊在BMPR-IB基因的相应位置上并未发生与Booroola Merino羊相同的突变,同时也未检测到BMP15的FecXI、FecXH、FecXB基因及GDF9的FecGH基因,因此排除了这5个突变位点存在控制福清山羊高繁殖力主效基因的可能性。     

BMPR-IB、BMP15和GDF9基因多态性与崂山奶山羊产羔数的相关研究

为了研究骨形态发生蛋白受体(BMPR-IB)、骨形态发生蛋白15(BMP15)、生长分化因子(GDF9)基因是否与崂山奶山羊的产羔数相关,试验采用PCR-RFLP法对290只经产崂山奶山羊BMPR-IB基因的FecB突变、BMP15基因的FecXH和FecXI突变、GDF9基因的G8突变(GDF9基因编码区1 184 bp处的碱基突变C→T)进行多态性检测,并探索与产羔数的关系。结果表明:崂山奶山羊BMPR-IB、BMP15、GDF9基因均只有1种基因型,不存在多态性。因此,BMPR-IB、BMP15、GDF9基因不能作为崂山奶山羊产羔数(多胎性状)的候选基因。     

湖羊BMPR-IB、BMP15和GDF9基因的RFLP分析

以BMPR-IB、BMP15、GDF9基因作为湖羊多胎性状候选基因,采用PCR-RFLP技术检测了53只湖羊上述候选基因的单核苷酸多态性。结果表明:湖羊BMPR-IB基因的FecB位点只存在BB和+B两种基因型,二者的基因型频率分别为0.981 1和0.018 9;B等位基因为绝对优势等位基因,基因频率为0.990 6。未检测到BMP15基因的B4(FecXB)突变和GDF9基因的G8(FecGH)突变。因此,推测BMPR-IB基因的FecB位点是湖羊多胎性的主效基因,而BMP15基因和GDF9基因与湖羊群产羔数关系不大。研究结果同时反映了所测湖羊群体是一个高度纯化的宝贵绵羊品种资源。     

云南黑山羊新品种和龙陵黄山羊高繁基因的研究

为了分析云南黑山羊新品种和龙陵黄山羊中骨形态发生蛋白受体IB(BMPR-IB)、骨形态发生蛋白15(BMP15)和生长分化因子9(GDF9)基因的多态性与产羔数的关系,采用PCR-SSCP与测序相结合的技术检测204个山羊样品的多态性,同时研究这3个基因对山羊繁殖力的影响。结果显示:龙陵黄山羊和云南黑山羊新品种中并未检测到FecB、FecX~I、FecX~H、FecX~L、FecX~G、FecX~B、FecG~H位点上的突变,碱基序列分析证实BMP15基因编码序列808位上C→G的突变,导致了氨基酸序列第270位由谷氨酰胺到谷氨酸的改变;云南黑山羊新品种和龙陵黄山羊在该位点都检测到了AA、AB和BB 3种基因型,基因频率分别为0.785/0.169/0.046和0.514/0.324/0.162,不同基因型的山羊产羔数最小二乘均值之间无显著差异(P0.05),表明BMPR-IB、BMP15和GDF9基因中8个突变位点对云南黑山羊新品种和龙陵黄山羊产羔数并无显著影响。     

哈萨克羊GDF9、ESR、BMPR-IB基因的PCR-RFLP多态性研究

为了探索绵羊多胎性的分子机理以及为绵羊繁殖力的标记辅助选择和育种提供理论依据,特开展了哈萨克羊品种多胎主效基因筛选和标记辅助育种研究。实验采用PCR-RFLP方法,对哈萨克羊GDF9、ESR、BMPR-IB基因进行PCR-RFLP多态性检测。结果表明:在哈萨克羊群体中GDF9、ESR、BMPR-IB基因的酶切位点上均未发现多态性,说明GDF9、ESR、BMPR-IB基因的酶切位点均不能作为控制哈萨克羊繁殖力的潜在分子标记位点。     

戴云山羊高繁殖力候选基因BMPR-IB、BMP15、GDF9的研究

为了初步探索戴云山羊高繁殖力遗传分子机制,采用PCR直接测序法检测BMPR-IB、BMP15、GDF9基因在戴云山羊群体中的单核苷酸多态性.结果表明:在GDF9基因内含子区域发生了2处点突变(2471A→T,884bp处插入T碱基),但均未引起氨基酸的改变;BMP15基因1176处发现3个碱基(TGG)的插入,导致280位一个Gly(甘氨酸)的插入,一处A→T突变(227位Ser丝氨酸→Thr苏氨酸的改变);BMPR-IB基因外显子区域未发生突变.     

山羊生长分化因子9基因FecTT突变检测

本研究采用PCR-RFLP方法检测生长分化因子9(growth differentiation factor 9, GDF9)基因FecTT突变在济宁青山羊、贵州白山羊、大足黑山羊、陕南白山羊、古蔺马羊、马头山羊、波尔山羊、成都麻羊、萨能奶山羊、文登奶山羊、板角山羊、金堂黑山羊、关中奶山羊、川东白山羊、安哥拉山羊、辽宁绒山羊、内蒙古绒山羊等17个山羊品种中的分布。结果表明,在17个山羊品种中都未检测到FecTT突变,提示,GDF9基因影响Thoka绵羊高繁殖力的FecTT突变对以上17个山羊品种的繁殖力均没有显著影响。     

6个绵羊群体BMP15、GDF9基因多态性分析

本研究旨在检测新吉细毛羊、中国美利奴羊（无角型）、东北细毛羊、杜×寒杂交羊（F1）、杜×寒杂交羊（F3）、白头杜泊羊6个绵羊群体骨形态发生蛋白15（bone morphogenetic protein 15,BMP15）、生长分化因子9（growth differentiation factor 9,GDF9）基因的多态性,为绵羊繁殖力的标记辅助选择和育种提供理论依据。以6个绵羊群体共378只个体为研究对象,利用PCR-SSCP技术检测BMP15、GDF9基因多态性;用DNAStar软件与I-TASSER软件预测蛋白质二级结构和三级结构;计算基因频率、基因型频率、Hardy-Weinberg平衡、杂合度（He）、纯合度（Ho）、有效等位基因数（Ne）和多态信息含量（PIC）。运用GraphPad Prism 6软件对新吉细毛羊群体BMP15基因多态性与产羔数进行相关性分析。结果显示,BMP15基因P1引物扩增片段存在多态性,6个群体共呈现3种基因型：AA、AC、CC,该突变为BMP15基因外显子1上58-60 bp 3个碱基（CTT）缺失,新吉细毛羊、杜×寒杂交羊（F3）、白头杜泊羊的χ²值均达到显著水平,处于Hardy-Weinberg不平衡状态,不同基因型间新吉细毛羊平均产羔数差异均不显著（P>0.05）;GDF9基因P2引物扩增片段存在多态性,6个群体共呈现3种基因型：DD、DE、EE,其中新吉细毛羊仅有1种基因型：DD,该突变为GDF9基因外显子2上477 bp处T→C的转换,该突变未导致氨基酸的改变,除新吉细毛羊外其余5个群体χ²值均未达到显著水平,处于Hardy-Weinberg平衡状态,说明GDF9基因T477C突变不能作为新吉细毛羊多胎性状遗传标记位点。     

绵羊BMP15基因外显子单核苷酸多态性研究

以控制Belclare和 Cambridge绵羊高繁殖力的骨形态发生蛋白15( Bone morphogenetic protein 15,BMP15)基因为候选基因,采用PCR-SSCP技术检测BMP15基因在中国部分绵羊品种 (蒙古羊、内蒙古细毛羊、呼伦贝尔羊、中国美利奴羊)中的单核苷酸多态性,同时研究突变位点对绵羊繁殖力的影响.结果表明:在4个绵羊品种中均检测到BMP15的突变,检测到的BMP15突变在4个绵羊品种中均呈现3种基因型.这表明BMP15基因外显子Ⅰ中虽然存在突变位点,但这一突变位点对绵羊繁殖力高低没有显著影响.     

与绵羊多胎繁殖性状相关的BMP家族基因的研究进展

文章就BMP家族基因中与绵羊多胎繁殖性状相关的主效基因和候选基因进行了综述,主要包括BMP2、BMP4、BMP15、GDF9以及BMP的受体BMPRH、BMPR—IB等基因的最新研究进展,阐述了其对绵羊繁殖率的影响,供研究者参考。     

鲁中肉羊BMPR1B、BMP15和GDF9基因多态性与产羔数关联分析

为分析绵羊已知多羔主效基因BMPR1B、BMP15和GDF9的多态性与鲁中肉羊产羔数之间的关系,采用Sequenom MassARRAY■SNP技术检测鲁中肉羊BMPR1B、BMP15和GDF9基因中已知主效位点多态性,并与产羔数进行关联分析。结果表明:BMPR1B基因在鲁中肉羊中存在FecB突变,GDF9基因6个分型位点仅4个位点(G1、G3、G4、G5)在鲁中肉羊中存在多态,BMP15基因中5个分型位点在鲁中肉羊中均不存在多态性。FecB基因3种基因型(CC、CT、TT)突变频率分别为0.16、0.58和0.26,此位点多态性较高(0.25相似文献   

Mutations in bone morphogenetic protein 15 and growth differentiation factor 9 genes are associated with increased litter size in fat-tailed sheep breeds

The objective of this study was to investigate association between GDF9 and BMP15 gene polymorphism and litter size in fat-tailed sheep, a total of 97 mature ewes from four breeds (Afshari=19; Baluchi=18; Makui=30 and Mehraban=30) were genotyped for the BMP15 HinfI and GDF9 HhaI polymorphisms by PCR-RFLP technique. The highest and lowest mutant allele frequencies were found in Makui (0.27) and Afshari (0.10) sheep for the BMP15 gene and in Afshari (0.24) and Mehraban (0.18) sheep for the GDF9 gene, respectively. Litter size was significantly influenced by genotype of the ewe for two genes (P < 0.01). Heterozygous genotypes for both loci showed higher litter size than homozygous genotypes (P < 0.01). None of the individuals carried homozygous genotype for both of the GDF9 and BMP15 variants in these breeds. The individuals carrying the mutant allele for one of the investigated candidate gene still showed fertile phenotype. Thus, existence of homozygosity at one of the BMP15 and GDF9 variant is not probably able to block normal hormonal pathway of reproduction in fat-tailed sheep.     

黔北麻羊生长分化因子9和骨形态发生蛋白15基因遗传变异分析

试验将生长分化因子9(GDF9)基因和骨形态发生蛋白15(BMP15)基因作为候选基因,采用直接测序法检测GDF9和BMP15基因在黔北麻羊中的单核苷酸多态性。结果表明,在GDF9基因外显子2的562 bp处发生了突变(A→C),导致谷氨酰胺突变为脯氨酸,检测到AA、Aa 2种基因型,基因型频率分别为0.7273、0.2727;A、a等位基因频率分别为0.8637、0.1363。BMP15基因外显子2的480 bp处发生了突变(C→G),导致谷氨酰胺突变为谷氨酸,检测到BB、Bb 2种基因型,基因型频率分别为0.7000、0.3000;B、b等位基因频率分别为0.8500、0.1500。     

Genetic Aspects of Reproduction in Sheep

Maintenance of high levels of realized fertility (defined as the percentage of ewes that lamb) and appropriate levels of fecundity are critical for efficient sheep production. The optimal level of fecundity in most situations is well below the maximum attainable level and can be targeted by combining selection among and within breeds with use of an expanding array of single-gene mutations affecting ovulation rate and litter size. The heritability of litter size is approximately 0.10, allowing changes of up to 2%/year from simple mass selection. Mutations in several genes associated with the transforming growth factor β superfamily ( BMPRIB , GDF9 and sex-linked BMP15 ) can increase ovulation rates by 0.7–1.5 ova in heterozygous ewes. However, ewes that are homozygous for BMP15 or GDF9 mutations are sterile, so use of these mutations requires carefully structured breeding programmes. Improvements in fertility may be critical for autumn lambing or programmes that aspire to lamb throughout the year. Selection to improve fertility in spring matings has been successful; selected adult ewes have lambing rates of 80–85% in October and early November. The selected ewes have a dramatically reduced seasonal anestrus, and many ewes continue to cycle during spring and summer. Major genes affecting seasonal breeding have not been identified in sheep. Polymorphisms in the melatonin receptor 1a gene appear to be associated with seasonal breeding in some, but not all breeds. However, functional genomic studies of genes associated with circadian and circannual rhythms have potential to reveal additional candidate genes involved in seasonal breeding.     

Effects of Growth Differentiation Factor 9 and Bone Morphogenetic Protein 15 on the in vitro Maturation of Porcine Oocytes

Growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) are members of the transforming growth factor‐β (TGF‐β) family, and their roles in oocyte maturation and cumulus expansion are well known in the mouse and human, but not in the pig. We investigated GDF9 and BMP15 expressions in porcine oocytes during in vitro maturation. A significant increase in the mRNA levels of GDF9 and BMP15 was observed at germinal vesicle breakdown, with expression levels peaking at metaphase I (MI), but decreasing at metaphase II (MII). GDF9 and BMP15 protein localized to the oocyte cytoplasm. While treatment with GDF9 and BMP15 increased the expression of genes involved in both oocyte maturation (c‐mos, cyclinb1 and cdc2) and cumulus expansion (has2, ptgs2, ptx3 and tnfaip6), SB431542 (a TGFβ–GDF9 inhibitor) decreased meiotic maturation at MII. Following parthenogenetic activation, the percentage of blastocysts in SB431542 treatment was lower than in the control (41.3% and 74.4%, respectively). Treatment with GDF9 and BMP15 also increased the mRNA levels of maternal genes such as c‐mos [a regulatory subunit of mitogen‐activated protein kinase (MAPK)], and cyclinb1 and cdc2 [regulatory subunits of maturation/M‐phase‐promoting factor (MPF)]; however, SB431542 significantly decreased their mRNA levels. These data were supported by poly (A)‐test PCR and protein activity analyses. Our results show that GDF9 and BMP15 participate in cumulus expansion and that they stimulate MPF and MAPK activities in porcine oocytes during in vitro maturation.