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西农萨能奶山羊CSN1S2基因多态与产奶量、体尺指标的相关分析 总被引：4，自引：0，他引：4
利用PCR2RFLP技术对69只西农萨能奶山羊的CSN1S2基因进行多态性分析。扩增产物为310bp的CSN1S2F基因片段,被Alw26I限制性内切酶消化后表现多态性。F等位基因为310bp,N等位基因为179和131bp,相应地,FF基因型为310bp,NF基因型为310,179和131bp,NN基因型为179和131bp。在该群体中,F等位基因频率为010870,N等位基因频率为019130,处于Hardy2Weinberg平衡状态。将西农萨能奶山羊CSN1S2F基因座不同基因型与平均产奶量进行相关分析,结果表明:CSN1S2的不同基因型对平均产奶量有显著影响,NN基因型个体产奶量最高,FF基因型个体产奶量最低,且FF基因型个体平均产奶量显著低于NN基因型个体(P<0105)。将西农萨能奶山羊CSN1S2F基因座不同基因型与体尺指标进行相关分析,结果表明:在初生重和体高指标上,NF基因型个体显著优于NN基因型个体(P<0105);在体斜长和管围指标上,NF基因型个体略优于NN基因型个体,但差异不显著(P>0105);在成年体重和胸围指标上,不同的基因型个体间无差异。 相似文献
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本研究旨在克隆牦牛酪蛋白基因家族(CSN1S1、CSN1S2、CSN2和CSN3)的CDS区序列,鉴定其在牦牛不同组织中的表达水平。选取4岁龄左右处于泌乳期的健康类乌齐母牦牛3头,屠宰后分别采集乳腺、心脏、肝脏、骨骼肌组织,分别提取组织总RNA并反转录为cDNA,设计酪蛋白基因家族特异性引物扩增酪蛋白基因家族序列,进行生物信息学分析,并利用实时荧光定量PCR法分别检测酪蛋白家族基因mRNA水平。结果显示,克隆得到CSN1S1、CSN1S2、CSN2和CSN3基因cDNA序列分别为919、832、805和715bp,其CDS区全长分别为645、669、690和585bp,分别编码214、222、259和194个氨基酸残基。类乌齐牦牛酪蛋白基因家族与黄牛亲缘关系最近,其次是印度水牛,而与单胃动物猪的亲缘关系最远。组织表达结果显示,酪蛋白基因家族在组织中广泛表达,其中在乳腺组织中的表达量最高,其次是骨骼肌组织。在乳腺组织中CSN1S1、CSN1S2、CSN2基因之间表达量差异不显著(P>0.05),但CSN2基因表达量显著高于CSN3基因(P<0.05)。以上结果为酪蛋白基因家族在牦牛乳腺蛋白质代谢调控机制的研究提供了参考依据。 相似文献
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应用Real-time PCR方法检测不同浓度(0、1、10和100 ng/mL)胰岛素样生长因子-Ⅰ(IGF-Ⅰ)处理后的体外培养奶牛乳腺上皮细胞乳蛋白和脂肪合成相关基因mRNA的相对表达量。结果表明,添加不同浓度IGF-Ⅰ对体外培养的奶牛乳腺上皮细胞IGF-Ⅰ受体基因(IGFIR)、IGF-Ⅰ结合蛋白-3基因(IGFBP3)、α-s1-酪蛋白基因(CSN1S1)和κ-酪蛋白基因(CSN3)mRNA的相对表达丰度均无显著影响(P>0.05)。随着IGF-Ⅰ添加浓度的增加,β-酪蛋白基因(CSN2)、乙酰辅酶A羧化酶基因(ACACA)、脂肪酸合成酶基因(FASN)和脂肪酸结合蛋白-3基因(FABP3)mRNA的相对表达丰度显著上调(P<0.05)。提示,IGF-Ⅰ作为一种重要细胞因子参与调节乳腺上皮细胞乳蛋白和乳脂肪相关基因mRNA的表达。 相似文献
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含蛋氨酸二肽影响奶牛乳腺上皮细胞乳蛋白合成相关基因表达 总被引：1，自引：0，他引：1
本试验旨在研究含蛋氨酸(Met)二肽对奶牛乳腺上皮细胞(BMECs)内乳蛋白合成相关基因表达的影响。试验分3部分,均采用单因子完全随机试验设计,Met的添加浓度及培养时间分别为60μg/m L(0.402 mmol/L)、48 h。第1部分,培养液添加8种含Met二肽[蛋氨酸-蛋氨酸(P-Met-Met)、蛋氨酸-赖氨酸(P-Met-Lys)、蛋氨酸-色氨酸(P-Met-Trp)、蛋氨酸-苯丙氨酸(P-Met-Phe)、蛋氨酸-苏氨酸(P-Met-Thr)、蛋氨酸-异亮氨酸(P-Met-Ile)、蛋氨酸-亮氨酸(P-Met-Leu)、蛋氨酸-缬氨酸(P-Met-Val)],以不添加二肽为对照,测定BMECs乳蛋白合成相关基因(αs1-酪蛋白、β-酪蛋白、κ-酪蛋白、β-乳球蛋白、Ⅱ型小肽转运载体和氨肽酶氮)的表达量;第2部分,培养液添加8种与上述二肽对应的游离氨基酸(F-Met-Met、F-Met-Lys、F-MetTrp、F-M et-Phe、F-M et-Thr、F-M et-Ile、F-M et-Leu、F-M et-Val),以不添加游离氨基酸为对照,测定BM ECs乳蛋白合成相关基因的表达量;第3部分,用二肽等物质的量替代相应游离氨基酸,测定BMECs乳蛋白合成相关基因的表达量以及细胞内外氨肽酶含量。结果表明:P-Met-Met和P-M et-Lys组较对照组和其他二肽组上调了αs1-酪蛋白和β-酪蛋白基因的表达量,且P-M et-M et组优于P-Met-Lys组。F-Met-Met和F-Met-Lys组较对照组和其他游离氨基酸组显著提高了αs1-酪蛋白基因的表达量(P0.05)。除P-Met-Val和P-Met-Leu组外,其他二肽替代游离氨基酸后均不同程度地提高了乳蛋白和Ⅱ型小肽转运载体基因的表达量,其中P-Met-Met表现出较好的促进效果。总之,含Met二肽等量替代对应的游离氨基酸能够促进乳蛋白基因的表达,其中尤以P-Met-Met的效果最好。 相似文献
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María Gabriela Pizarro Inostroza Vincenzo Landi Francisco Javier Navas González José Manuel León Jurado María del Amparo Martínez Martínez Javier Fernández Álvarez Juan Vicente Delgado Bermejo 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2020,137(4):407-422
Goat milk casein proteins (αS1, αS2, β and κ) are encoded by four loci (CSN1S1, CSN1S2, CSN2 and CSN3, respectively) clustered within 250 kb in chromosome 6. In this study, 159 Murciano-Granadina goats were genotyped for 48 SNPs within the entire casein region. Phenotypes on milk yield and components were obtained from 2,594 dairy registries. Additive and dominance effects on milk composition and quality were studied using non-parametric tests and principal component analysis to prevent SNPs multicollinearity. Two deletions in exon 4 (CSN1S1 and CSN3), one in exon 7 (CSN2) and one in exon 15 (CSN1S2) have been found at frequencies ranging from 0.12 to 0.50. Bonferroni-corrected significant SNP additive and dominance effects were found for milk yield, fat, protein, dry matter and lactose, and somatic cells. Exons 15 and 7 were significantly associated with milk yield and components except for lactose and somatic cells, while exon 4 was significantly associated with milk yield and components except for protein and dry matter. SNPs' associations with somatic cells were less frequent and weaker than those with milk yield and components. As caseins increase, somatic cells decrease, reducing milk enzymatic activity and consumption suitability. Hence, including molecular information in breeding schemes may promote production efficiency, as selecting against undesirable alleles could prevent the compromises derived from their dominance effects. 相似文献
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应用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和体外模拟消化研究羊乳蛋白质组成和消化特性。结果表明：羊乳蛋白质主要由酪蛋白和乳清蛋白组成,酪蛋白主要由αs1-酪蛋白、αs2-酪蛋白、β-酪蛋白和κ-酪蛋白组成,在酪蛋白中的相对含量分别为23.10%、30.39%、38.09%和8.42%;乳清蛋白主要由α-乳白蛋白、β-乳球蛋白、乳铁蛋白、血清白蛋白和免疫球蛋白组成,在乳清蛋白中的相对含量分别为24.59%、57.50%、4.35%、8.69%和4.88%;羊乳酪蛋白主要在肠液中消化,在胃液中消化120 min时 相似文献
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牛乳含有丰富的蛋白质、脂肪等营养素，是婴幼儿重要的营养来源，牛乳蛋白也是诱发婴幼儿食物过敏的
主要因素之一。对5 种市售婴幼儿乳粉进行体外模拟静态消化，通过电泳、水解度测定分析蛋白消化稳定性，并利
用间接竞争酶联免疫吸附实验评价乳清蛋白的抗原性。结果表明：5 种配方乳粉中，α-乳白蛋白和β-乳球蛋白在样
品1、2、4中具有较好的消化稳定性，胃肠消化后，样品1中α-乳白蛋白和β-乳球蛋白的抗原性降低，样品2中2 种过
敏原蛋白的抗原性增加，样品4中α-乳白蛋白的抗原性增加，β-乳球蛋白的抗原性降低；α-乳白蛋白和β-乳球蛋白在
样品3和5中消化稳定性较低，经胃肠消化后，样品5中2 种过敏原蛋白的抗原性增加，样品3中α-乳白蛋白的抗原性
增加，β-乳球蛋白的抗原性降低。 相似文献
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【目的】 通过分析关中奶山羊αs1-酪蛋白（alpha-s1 casein，CSN1S1）基因组织表达谱及其生物信息学功能，初步探究CSN1S1基因在奶山羊乳成分合成中发挥的作用。【方法】 以关中奶山羊为研究对象，利用PCR扩增并克隆CSNIS1-1和CSN1S1-2 2个突变形态，并利用ProtParam、NetPhos、SingalP 4.1 Server、NPS和Phyre2等多种生物信息软件和在线工具对CSN1S1基因及其突变形态的蛋白质结构、理化性质、磷酸化位点等进行分析，通过实时荧光定量PCR检测CSNIS1-1和CSN1S1-2在关中奶山羊肝脏、脾脏、乳腺、肾脏、子宫、输卵管6个组织中的相对表达量。【结果】 关中奶山羊乳腺上皮细胞中存在CSN1S1-1和CSN1S1-2 2种突变形态。对测序结果比对显示，CSN1S1-1存在3个碱基的突变，CSN1S1-2不仅存在3个碱基的突变，还存在6个碱基的缺失。CSN1S1-1与CSN1S1蛋白相似性为99.07%，CSN1S1-2与CSN1S1蛋白相似性为97.20%。蛋白理化性质分析显示，CSN1S1-1中碱基的突变导致第31位亮氨酸变成脯氨酸、第92位谷氨酰胺变成谷氨酸；CSN1S1-2除上述改变之外，第83位由丝氨酸变成半胱氨酸、第84位由谷氨酸变成谷氨酰胺，还存在第81和82位2个丝氨酸的缺失。实时荧光定量PCR结果显示，CSN1S1-1和CSN1S1-2在关中奶山羊各组织中相对表达趋势基本相同，其中在乳腺、子宫、输卵管中相对表达量较高，在肝脏、脾脏和肾脏中基本不表达，在子宫中CSN1S1-2的表达量显著高于CSN1S1-1（P<0.05）。【结论】 关中奶山羊乳腺上皮细胞中存在CSN1S1-1和CSN1S1-2 2种突变形态，且2种突变形态与CSN1S1的蛋白相似性均达到97%以上。CSN1S1-1在蛋白结构上与CSN1S1已有明显区别，而CSN1S1-2中6个碱基的缺失是导致氨基酸改变、磷酸化位点缺失与移位的直接原因。CSN1S1-2在子宫中表达显著高于CSN1S1-1，其可能在子宫中发挥潜在功能，还有待进一步探索。 相似文献
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Male germ cells modified by foreign genes can be used to generate transgenic chicken. In this study, in vivo transfection of chicken testis with an EGFP‐LacZ dual reporter expression vector was performed. Large‐scale plasmid DNA preparation of the EGFP‐LacZ eukaryotic expression vector was carried out and efficient transfection of chicken testicle cells using the prepared plasmid DNA was confirmed in vitro. The reporter plasmid was directly injected into adult rooster testes. Semen samples were collected on 10‐days post‐transfection and every other day thereafter; and a total of six collections were made. Semen slides were subjected to fluorescence microscopy and β‐galactosidase activity assay to identify sperms carrying the reporter genes. The presence of EGFP and LacZ was further confirmed by PCR amplification with sperm genomic DNA as template. The testicles of those birds were subjected to cryostat sectioning, fluorescence microscopy and β‐galactosidase activity assay. The results showed that sperms with green fluorescence were not observed on semen slides; however, sperms positive for β‐galactosidase were detected. Specific amplicons of EGFP and lacZ were detected in four of the six sequentially collected semen samples. Fluorescence microscopy of the corresponding semen slides revealed yellow‐green fluorescence, but not clear green fluorescence. The β‐galactosidase activity assay and GFP histochemistry using monoclonal antibodies demonstrated positive staining for subsets of testicle cells. Together, these results showed that direct injection of the dual reporter vector into adult rooster testis allowed in vivo transfection of chicken sperm precursor cells, which further developed into sperm containing EGFP‐LacZ. 相似文献
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Manjarin R Zamora V Wu G Steibel JP Kirkwood RN Taylor NP Wils-Plotz E Trifilo K Trottier NL 《Journal of animal science》2012,90(9):3088-3100
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D. Marletta S. Bordonaro A. M. Guastella G. D'Urso 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2004,121(1):52-56
In this study we surveyed two endangered Sicilian goat breeds (Girgentana and Argentata dell'Etna) for genetic polymorphism at the CSN1S2 locus. In a total of 537 goats, we detected CSN1S2A, CSN1S2B, CSN1S2C° (CSN1S2C + CSN1S2E), CSN1S2D, CSN1S2F and CSN1S2O alleles by means of various polymerase chain reaction (PCR), allele specific‐PCR (AS–PCR) and PCR‐restriction fragment length polymorphism (RFLP) reactions with the aim of improving the knowledge of the genetic resource of these two breeds. Three and five alleles, with six and twelve genotypes, were identified at CSN1S2 locus, in Girgentana and Argentata, respectively. Argentata dell'Etna showed a higher degree of genetic variation. The allelic and genotypic distribution seems to be significantly different (p < 0.001) in the two breeds. In Argentata the rare null allele (CSN1S2O) was found at low frequency (0.033); this genetic peculiarity makes its preservation worthwhile. 相似文献
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Ikawa K Aoki M Ichikawa M Itagaki T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2011,73(10):1359-1361
In the present study, we tried to detect DNA for ribosomal RNA genes of piroplasma parasites from the liver or blood of 43 Japanese serows (Capricornis crispus) in Iwate Prefecture of Japan by polymerase chain reaction. Approximately 500-bp amplicons were obtained in 35 (81.4%) of the 43 samples by amplification for V4 hyper-variable regions of the 18S rRNA gene, and the amplicons were considered to be DNA of Theileria species. The complete nucleotide sequences (1,700 or 1709 bp) of the 18S rRNA gene were determined in 20 samples and were divided into 5 genotypes that were phylogenetically separated into two different lineages showing a polyphyletic relation. The Theileria DNAs of the two different lineages were considered to be those of distinct species. 相似文献
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Samer Alasaad Joerns Fickel Luca Rossi Mathieu Sarasa Buenaventura Ben?-tez-Camacho José E Granados Ramón C Soriguer 《Acta veterinaria Scandinavica》2012,54(1):56
Background
Hybridization between closely related wild and domestic species is of great concern because it can alter the evolutionary integrity of the affected populations. The high allelic variability of Major Histocompatibility Complex (MHC) loci usually excludes them from being used in studies to detect hybridization events. However, if a) the parental species don’t share alleles, and b) one of the parental species possesses an exceptionally low number of alleles (to facilitate analysis), then even MHC loci have the potential to detect hybrids.Results
By genotyping the exon2 of the MHC class II DRB1 locus, we were able to detect hybridization between domestic goats (Capra hircus) and free-ranging Iberian ibex (Capra pyrenaica hispanica) by molecular means.Conclusions
This is the first documentation of a Capra pyrenaica × Capra hircus hybridization, which presented us the opportunity to test the applicability of MHC loci as new, simple, cost-effective, and time-saving approach to detect hybridization between wild species and their domesticated relatives, thus adding value to MHC genes role in animal conservation and management. 相似文献16.
本试验旨在研究不同蛋白质源饲料中添加α-酮戊二酸对杂交鲟(Acipenser schrenckii♀×Acipenser baeri)幼鱼肝脏谷氨酰胺(Gln)含量、抗氧化能力及生长激素(GH)、胰岛素样生长因子-Ⅰ(IGF-Ⅰ)基因表达的影响。分别以大豆浓缩蛋白和大豆浓缩蛋白+鱼粉为蛋白质源,设2个α-酮戊二酸(AKG)添加量(0和1%),配制4种试验饲料。选取平均体重为(7.65±0.04)g的杂交鲟幼鱼500尾,随机分为4组,每组5个重复,每个重复25尾,养殖周期为8周。结果表明:饲料中添加1%AKG显著提高肝脏Gln含量和谷氨酰氨合成酶(GS)活性(P0.05),对肝脏碱性磷酸酶(ALP)活性无显著影响(P0.05)。蛋白质源对肝脏Gln含量、GS活性和ALP活性无显著影响(P0.05),且蛋白质源和AKG添加量对肝脏Gln含量、GS活性和ALP活性无显著交互作用(P0.05)。饲料中添加1%AKG显著降低肝脏丙二醛(MDA)含量,显著提高肝脏还原型谷胱甘肽(GSH)含量(P0.05)。与大豆浓缩蛋白+鱼粉作为蛋白质源相比,大豆浓缩蛋白作为蛋白质源显著提高肝脏GSH含量(P0.05)。蛋白质源和AKG添加量对肝脏超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性均无显著影响(P0.05),且蛋白质源和AKG添加量对肝脏各抗氧化指标无显著交互作用(P0.05)。饲料中添加1%AKG显著提高肝脏GH、IGF-Ⅰ基因的相对表达量(P0.05)。与大豆浓缩蛋白+鱼粉作为蛋白质源相比,大豆浓缩蛋白作为蛋白质源显著提高肝脏IGF-Ⅰ和GH基因的相对表达量(P0.05)。蛋白质源和AKG添加量对肝脏GH、IGF-Ⅰ基因的相对表达量无显著交互作用(P0.05)。综上所述,杂交鲟幼鱼饲料中添加1%AKG可以通过提高肝脏中GS的活性进而提高Gln的含量,通过提高肝脏中GSH的含量进而降低MDA的含量,并可提高肝脏中生长相关基因GH、IGF-Ⅰ的表达。 相似文献
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The MHC of cattle, known as the bovine leukocyte antigen (BoLA) complex, plays an integral role in disease and parasite susceptibility, and immune responsiveness of the host. While susceptibility to tick infestation in cattle is believed to be heritable, genes that may be responsible for the manifestation of this phenotype remain elusive. In an effort to analyze the role that genes within the BoLA complex may play in host resistance to ticks, we have evaluated components of this system within a herd of cattle established at our laboratory that has been phenotyped for ectoparasite susceptibility. Of three microsatellite loci within the BoLA complex analyzed, alleles of two microsatellite loci within the BoLA class IIa cluster (DRB1-118 and DRB3-174) associated with the tick-resistant phenotype, prompting further investigation of gene sequences within the DRB3 region. DRB3 is a class IIa gene, the second exon of which is highly polymorphic since it encodes the antigen recognition site of the DR class II molecule. Analysis of the second exon of the DRB3 gene from the phenotyped calves in our herd revealed a significant association between the DRB3*4401 allele and the tick-resistant phenotype. To our knowledge, this is the first report of a putative association between a class IIa DRB3 sequence and host resistance to the Lone Star tick. Elucidation of the mechanism involved in tick resistance will contribute to improving breeding schemes for parasite resistance, which will be beneficial to the cattle industry. 相似文献
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The preliminary study on the effects of growth hormone and insulin‐like growth factor‐I on κ‐casein synthesis in bovine mammary epithelial cells in vitro 
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下载免费PDF全文 M. Z. Wang Y. Ji C. Wang L. M. Chen H. R. Wang J. J. Loor 《Journal of animal physiology and animal nutrition》2016,100(2):251-255
The effects of growth hormone (GH) and insulin‐like growth factor‐I (IGF‐I) on protein synthesis and gene expression of κ‐casein in bovine mammary epithelial cell in vitro were studied. The treatments were designed as follows: the growth medium without serum was set as the control group, while the treatments were medium supplemented with GH (100 ng/ml), IGF‐I (100 ng/ml), and GH (100 ng/ml) + IGF‐I (100 ng/ml). The quantity of κ‐casein protein was measured by ELISA, and the κ‐casein gene (CSN3) expression was examined by real‐time quantitative PCR (RT‐qPCR). Compared with the control group, all the experimental groups had greater (p < 0.05) expression of CSN3. The concentration of κ‐casein followed a similar response as CSN3, but the difference between the treatments and the control was not statistically significant (p > 0.05). Furthermore, no synergistic effect of GH and IGF‐I was observed for both the κ‐casein concentration and CSN3 expression. It is therefore concluded that GH or IGF‐I can independently promote the expression of CSN3 in bovine mammary epithelial cells in vitro. 相似文献
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Anna Wedholm Elin Hallén Lotte Bach Larsen Helena Lindmark-Månsson Anders Hans Karlsson Toomas Allmere 《Acta Agriculturae Scandinavica, Section A - Animal Sciences》2013,63(1):8-15
Abstract The milk composition in a Swedish herd, consisting of Swedish Red and White cows (SRB) and Swedish Holstein cows (SLB), and in a Danish herd, consisting of Danish Holstein-Friesian cows (SDM), was evaluated. Concentrations of the major milk proteins (κ-casein, αS1-casein, β-casein, β-lactoglobulin A, β-lactoglobulin B and α-lactalbumin) of 134 individual milk samples were determined by reversed phase (RP) HPLC. Other parameters determined included milk fat, urea, lactose, calcium, lactoferrin, somatic cell count and protein degradation (determined as level of free amino-terminals). Analysis of variance was used to compare concentrations of analysed milk variables between SRB and SLB or between SLB and SDM. Concentration of total protein, total casein, β-casein and κ-casein were significantly higher in SRB milk compared with SLB milk. Concentration of α-lactalbumin and calcium were significantly higher in SDM milk than in SLB milk. The concentration of urea was higher in SLB than in SDM milk and is suggested to reflect differences in feeding regimes between the investigated Swedish and Danish herd. 相似文献