中国荷斯坦牛POU1F1基因与PRL基因的多态性及其聚合效应对产奶性状的影响

文章采用DNA测序、PCR-RFLP和CRS-PCR技术对979头中国荷斯坦牛POU1F1基因与PRL基因进行研究,发现了3个新SNPs,分别是POU1F1基因第二外显子G1178C、PRL基因5侧翼区A906G和A1134G。采用SAS统计软件GLM程序,利用最小二乘法拟合线性模型,分析基因多态性与产奶性状的关系。结果表明:POU1F1基因1178位点GC基因型在产奶量、乳蛋白量、乳脂量方面均为优良基因型。PRL基因5侧翼区906位点AG基因型在产奶量方面为优良基因型,1134位点不同基因型产奶性状差异不显著。对PRL基因5侧翼区的906位点和POU1F1基因的1178位点进行基因互作分析,结果在乳脂率、乳蛋白率、产奶量、乳蛋白量和乳脂量方面各基因型组合之间均未观察到显著差异,说明基因聚合效应并不是单基因效应的简单相加,基因聚合效应在分子育种中具有更重要的意义。     

Association between polymorphisms of Myf5 and POU1F1 genes with growth and carcass traits in Hanwoo (Korean cattle)

Myogenic factor 5 (Myf5) and POU class 1 homeobox 1 (POU1F1) genes play important roles in growth and development of mammals. Bovine Myf5 and POU1F1 were characterized to detect genetic variation at these loci and to replace them to economic traits in 367 cattle representing Hanwoo (325) and Angus (37). Two single nucleotide polymorphisms (SNPs) were identified in intron 2 (A1948G SNP) of Myf5 and exon 6 (A15635G SNP) of POU1F1 by sequence analyses of genomic DNA. Statistical analysis indicated that the Myf5 polymorphisms significantly (0.05) associated backfat thickness and live weight at 6-months-of-age and that POU1F1 polymorphisms significantly influenced carcass weight and live weight at 24-month of age, and backfat thickness. The interaction between Myf5 and POU1F1 was significant on carcass weight, M. longissimus dorsi area, backfat thickness and marbling score. The results implicate Myf5 and POU1F1 as candidate genes of growth and carcass traits, and suggest that the interaction between Myf5 and POU1F1 strongly affect growth and carcass traits in cattle.     

猪POU1F1基因5’侧翼区克隆及序列分析

POU1F1基因是POU基因家族的成员之一,对哺乳动物的早期发育和相关基因启动表达起重要的调控作用。本文利用染色体步移技术,从长白猪基因组中首次克隆到了约1．5kb的POU1F1基因5’侧翼区序列,并利用相关软件对其进行了序列分析和物种间POU1F1基因5’侧翼区序列比对及进化树构建。结果表明所克隆的片段中含典型的TATA盒（-57）和类CAAT盒序列（-87）。TESS软件分析发现在启动子附近有一系列潜在的重要的转录因子结合位点。序列比对结果表明不同物种POU1F1基因的转录起始点上游-150bp区域保守性较强,可能是启动子的核心序列。其中猪与牛的同源性最高,其次是黑猩猩、人、狗,与大鼠、小鼠和鸡同源性较低,与斑马鱼序列同源性最低。同源序列主要集中在转录起始点上游-150bp至-1000bp区域。Vista中的MLAGAN程序构建的系统进化树真实反应了物种间进化关系。     

A TaqI PCR-RFLP Detecting a Novel SNP in Exon 2 of the Bovine POU1F1 Gene

PCR-SSCP and DNA sequencing methods were applied to reveal three novel single nucleotide polymorphisms (SNPs) in exon 2 of the POU1F1 gene in 963 Chinese cattle belonging to eight breeds. Among them, a silent SNP (NM_174579:c.545G > A) detected by TaqI endonuclease is described. Frequencies of the POU1F1-G allele varied from 0.685 to 1.000. The association of TaqI polymorphism with growth traits was analyzed in 251 Nanyang cattle. No significant associations of the TaqI polymorphism with body weight and average daily gain for different growth periods (6, 12, 18, and 24 months old) were observed (P > 0.05), as well as for body sizes (P > 0.05).     

POU1F1基因的遗传变异对南阳牛生长发育性状的影响

利用PCR-RFLP技术首次研究了南阳牛群体100个个体POU1F1基因多态性及其与体重、体尺等生长性状指标之间的相关性。结果表明,南阳牛群体POU1F1基因座的451bp的PCR产物被限制性酶Hinf Ⅰ消化后表现多态性,它们的等位基因A／B频率为：0．465／0．535,且处于Hardy-Weinberg平衡状态。同时,南阳牛群体POU1F1-Hinf Ⅰ基因座不同基因型与体重、体尺等生长性状指标相关分析的结果表明：南阳牛群体内BB与AB基因型个体在初生重、断奶前平均日增重、六月龄体重、体斜长和胸围以及十二月龄的体重、体高、体斜长和胸围指标上有显著差异,且BB〉AB（P〈0．05）;群体内BB型个体在十二月龄体重指标上显著高于群体的AA型个体,即BB〉AA（P〈0.05）,在其他各年龄段的各项体重和体尺指标上同样呈现出B等位基因高于A等位基因的一种趋势。初步认为BB基因型为优势基因型,相应地B为优势等位基因,对选择有正向效应,提示POU1F1基因的占等位基因可能与高生长发育性状有关。     

MC4R、POU1F1基因对京海黄鸡生长性能的遗传效应

以MC4R和POU1F1基因为候选基因, 采用PCR-SSCP和DNA测序技术检测两个候选基因在京海黄鸡群体中的单核苷酸多态性(SNPs), 同时对候选基因与京海黄鸡生长性能的相关性进行了研究。结果表明, MC4R基因编码区第662 bp位置有G→C碱基的点突变, 在京海黄鸡中检测到AA、AB、BB 3种基因型, A等位基因频率为0.929, B等位基因频率为0.071; 在POU1F1基因exon3在序列的第5 231 bp位置有一个A→T碱基的点突变, 检测到CC、CD、DD 3种基因型, C等位基因频率为0.500, D等位基因频率为0.500。采用GLM模型分析基因型对生长性能的遗传效应, 结果表明, MC4R基因AA基因型个体的4、8、12周龄体重显著地高于BB型个体(P<0.05), 16周龄体重差异极显著(P<0.01); POU1F1基因CD基因型个体体重极显著高于CC型和DD型(P<0.01)。因此推测MC4R和POU1F1基因可能是影响鸡生长性状的主效基因或与主效基因紧密连锁的标记基因, 能够在分子标记辅助选择中用于对鸡生长性状的遗传改良。     

Interactions of the bovine placental lactogen and prolactin receptor genes are associated with fertility traits in cattle

Decline in fertility of high-producing dairy cattle has become a global challenge to the dairy industry. Because of low heritability and complexity, it is difficult to find genetic markers for fertility traits in cattle. Here, we report the use of an in vitro fertilization (IVF) system and candidate gene approach to test genetic associations of a single-nucleotide polymorphism (SNP) in bovine placental lactogen (bPL), and its interactions with SNPs in the prolactin receptor (PRLR) and growth hormone receptor genes with fertility traits in an IVF system. The associations suggest a possible involvement of genetic interactions between bPL and PRLR in the fertilization and embryonic development processes, and the potential for application in a marker-assisted selection program.     

Genes of the RNASE5 pathway contain SNP associated with milk production traits in dairy cattle

Background

Identification of the processes and mutations responsible for the large genetic variation in milk production among dairy cattle has proved challenging. One approach is to identify a biological process potentially involved in milk production and to determine the genetic influence of all the genes included in the process or pathway. Angiogenin encoded by angiogenin, ribonuclease, RNase A family 5 (RNASE5) is relatively abundant in milk, and has been shown to regulate protein synthesis and act as a growth factor in epithelial cells in vitro. However, little is known about the role of angiogenin in the mammary gland or if the polymorphisms present in the bovine RNASE5 gene are associated with lactation and milk production traits in dairy cattle. Given the high economic value of increased protein in milk, we have tested the hypothesis that RNASE5 or genes in the RNASE5 pathway are associated with milk production traits. First, we constructed a “RNASE5 pathway” based on upstream and downstream interacting genes reported in the literature. We then tested SNP in close proximity to the genes of this pathway for association with milk production traits in a large dairy cattle dataset.

Results

The constructed RNASE5 pathway consisted of 11 genes. Association analysis between SNP in 1 Mb regions surrounding these genes and milk production traits revealed that more SNP than expected by chance were associated with milk protein percent (P < 0.05 significance). There was no significant association with other traits such as milk fat content or fertility.

Conclusions

These results support a role for the RNASE5 pathway in milk production, specifically milk protein percent, and indicate that polymorphisms in or near these genes explain a proportion of the variation for this trait. This method provides a novel way of understanding the underlying biology of lactation with implications for milk production and can be applied to any pathway or gene set to test whether they are responsible for the variation of complex traits.     

Polymorphisms and expression of the chicken POU1F1 gene associated with carcass traits

POU1F1 is an essential factor that regulates the development and reproduction of animal. The objective of the current research was to screen for polymorphism, expression of POU1F1 and their association with carcass quality traits. A total of 126 Erlang mountainous chickens from two strains (SD02 and SD03) were employed for testing. Seventeen single nucleotide polymorphisms (SNPs) were detected, but only two SNPs (g.96217999 T > C and g.96219442 C > T) were associated with carcass quality traits. In SD03 chicken, g.96217999 T > C genotypes were significantly associated with body weight (BW), carcass weight (CW), eviscerated weight (EW), and semi-eviscerated weight (SEW; P < 0.05), and was highly significantly associated with breast muscle weight (BMW) and abdominal fat weight (AW; P < 0.01). g.96219442 C > T was significantly associated with BW, EW, SEW (P < 0.05). However, these two SNPs were not significantly associated with any carcass traits in SD02 chicken. Diplotypes showed that in SD03 chicken, the haplotype [C: C] was the most favorable haplotype because it was associated with higher BW, CW, SEW, EW, BMW, and AW (P < 0.05). On the contrary, haplotype [T: T] was associated with lower carcass quality traits (P < 0.01). In addition, qRT-PCR revealed that at 13 weeks, the POU1F1 mRNA expression was significantly higher in breast muscle of cock compared to that of hens (P < 0.05), whereas there was no significant correlation between POU1F1 expression and carcass traits. These results suggested that POU1F1 could be a potential candidate gene for carcass traits in chicken.     

Single nucleotide polymorphisms in the growth hormone and insulin-like growth factor-1 genes are associated with milk production, body condition score and fertility traits in dairy cows

The somatotrophic axis (GH-IGF) is a key regulator of animal growth and development, affecting performance traits that include milk production, growth rate, body composition, and fertility. The aim of this study was to quantify the association of previously identified SNPs in bovine growth hormone (GH1) and insulin-like growth factor 1 (IGF-1) genes with direct performance trait measurements of lactation and fertility in Holstein-Friesian lactating dairy cows. Sixteen SNPs in both IGF-1 and GH1 were genotyped across 610 cows and association analyses were carried out with traits of economic importance including calving interval, pregnancy rate to first service and 305-day milk production, using animal linear mixed models accounting for additive genetic effects. Two IGF-1 SNPs, IGF1i1 and IGF1i2, were significantly associated with body condition score at calving, while a single IGF-1 SNP, IGF1i3, was significantly associated with milk production, including milk yield (means ± SEM; 751.3 ± 262.0 kg), fat yield (21.3 ± 10.2 kg) and protein yield (16.5 ± 8.0 kg) per lactation. Only one GH1 SNP, GH33, was significantly associated with milk protein yield in the second lactation (allele substitution effect of 9.8 ± 5.0 kg). Several GH1 SNPs were significantly associated with fertility, including GH32, GH35 and GH38 with calving to third parity (22.4 ± 11.3 days) (GH32 and GH38 only), pregnancy rate to first service (0.1%) and overall pregnancy rate (0.05%). The results of this study demonstrate the effects of variants of the somatotrophic axis on milk production and fertility traits in commercial dairy cattle.