秦川肉牛FABP3及FABP4基因SNP与肉质性状的关联性
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摘要
【目的】探索脂肪酸结合蛋白3(Fat acid binding proteins 3,FABP3)及脂肪酸结合蛋白4(FABP4)基因对秦川牛肉用新品系肉用性状的影响,为秦川肉牛的选育提供理论基础。【方法】随机选择18~24月龄健康的秦川肉牛571头,尾静脉采血10mL/头,提取血液DNA,PCR扩增FABP3及FABP4基因,检测其单核苷酸多态性(SNP)位点,并分析SNP位点基因型和单倍型组合对肉牛肉用性状(背膘厚、眼肌面积、肌间脂肪含量)的影响。【结果】测序发现,在FABP3基因第3个内含子上检测出1个SNP位点:g.60298C>T。在FABP4基因第1个内含子上检测出2个SNP位点:g.2686A>T和g.2834C>G;在第4外显子上检测出1个SNP位点:g.4220A>G。FABP3基因g.60298C>T位点不同基因型的肌内脂肪含量差异显著(P<0.05),TT基因型显著高于CT和CC基因型。对FABP4基因而言,g.2834C>G位点上的CC基因型肉用性状表现较优,其眼肌面积极显著高于GG和CT基因型(P<0.01),肌内脂肪含量显著高于GG和CG基因型(P<0.05);g.4220A>G位点上的TT基因型的肉用性状较优,其背膘厚显著高于CC和CT基因型(P<0.05)。H1H1是最优秀的单倍型组合,其眼肌面积和肌内脂肪含量分别极显著高于单倍型组合H3H3和H3H7(P<0.01)。【结论】FABP3及FABP4基因与秦川牛肉用新品系肉用性状关联紧密。
【Objective】In this study,the effects of FABP3 and FABP4 genetic variations on meat traits of Qinchuan beef cattle were evaluated to provide basis for breeding of Qinchuan beef cattle.【Method】A total of 571 Qinchuan beef cattle at the age of 18-24 months were chosen and blood samples with 10 ml were collected at the tail vein.Then,we extracted DNA of blood and the genes were cloned by PCR.The SNPs of FABP3 and FABP4 genes were amplified and effects of haplotypes and combined genotypes on meat traits(backfat thickness,ultrasound lean muscle area and intramuscular fat)were analyzed.【Result】There was one SNP(g.60298 C>T)in FABP3 third intron,while there were three SNPs sites in FABP4 totally,located at g.2686 A>T and g.2834 C>G of first intron and g.4220 A>G of fourth exon.Single nucleotide polymorphism(g.60298 C>T)of FABP3 was positively correlated with intramuscular fat(P<0.05),and the individuals with TT homozygotes had significantly higher IMF than those with CT genotype and CC genotype(P<0.01).At g.2834 C>G locus of FABP4,the individuals with CC homozygotes had higher ultrasound lean muscle area than GG and CG genotype significantly(P<0.05).At g.4220 A>G locus,the individuals with TT homozygotes had higher backfat thickness than CC and CT genotype significantly(P<0.05).H1H1(ACA-ACA)diplo type had better performance than H3H3(AGA-AGA)in ULA(P<0.01)and H3H7(AGA-TGA)in IMF(P<0.01).H1H1 was the most superior combined genotype.【Conclusion】SNPs mutations in FABP3 gene and FABP4 had significant impacts on meat traits of Qinchuan beef cattle.
【Objective】In this study,the effects of FABP3 and FABP4 genetic variations on meat traits of Qinchuan beef cattle were evaluated to provide basis for breeding of Qinchuan beef cattle.【Method】A total of 571 Qinchuan beef cattle at the age of 18-24 months were chosen and blood samples with 10 ml were collected at the tail vein.Then,we extracted DNA of blood and the genes were cloned by PCR.The SNPs of FABP3 and FABP4 genes were amplified and effects of haplotypes and combined genotypes on meat traits(backfat thickness,ultrasound lean muscle area and intramuscular fat)were analyzed.【Result】There was one SNP(g.60298 C>T)in FABP3 third intron,while there were three SNPs sites in FABP4 totally,located at g.2686 A>T and g.2834 C>G of first intron and g.4220 A>G of fourth exon.Single nucleotide polymorphism(g.60298 C>T)of FABP3 was positively correlated with intramuscular fat(P<0.05),and the individuals with TT homozygotes had significantly higher IMF than those with CT genotype and CC genotype(P<0.01).At g.2834 C>G locus of FABP4,the individuals with CC homozygotes had higher ultrasound lean muscle area than GG and CG genotype significantly(P<0.05).At g.4220 A>G locus,the individuals with TT homozygotes had higher backfat thickness than CC and CT genotype significantly(P<0.05).H1H1(ACA-ACA)diplo type had better performance than H3H3(AGA-AGA)in ULA(P<0.01)and H3H7(AGA-TGA)in IMF(P<0.01).H1H1 was the most superior combined genotype.【Conclusion】SNPs mutations in FABP3 gene and FABP4 had significant impacts on meat traits of Qinchuan beef cattle.
引文
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[16]李娟,杜新华,陈燕,等.牛FABP3转基因小鼠的遗传及表达特性研究[J].畜牧兽医学报,2015,46(10):1750-1758.Li J,Du X H,Chen Y,et al.Study on genetic characteristic and expression of bovine FABP3gene in transgenic mice[J].Chinese Journal of Animal and Veterinary Sciences,2015,46(10):1750-1758.
[17]Ye M H,Chen J L,Zhao G P,et al.Associations of A-FABP and H-FABP markers with the contnt of intramuscular fat in Beijing-You chicken[J].Anim Biotechnol,2010,21(1):14-24.
[18]Cho K H,Kim M J,Jeon G J,et al.Association of genetic variants for FABP3gene with back-fat thickness and intramuscular fat content in pig[J].Molecular Biology Reports,2011,38:2161-2166.
[19]Li X P,Kim S W,Choi J S,et al.Investigation of porcine FABP3and LEPR gene polymorphisms and mRNA expression for variation in intramuscular fat content[J].Molecular Biology Reports,2010,37:3931-3939.
[20]Blecha I M Z,Siqueira F,Ferreira A B R,et al.Identification and evaluation of polymorphisms in FABP3and FABP4in beef cattle[J].Genetics and Molecular Research,2015,14(4):16353-16363.
[21]Zhang J,Yang J H,Huang Y L,et al.Effects of different diet forms on growth performance and nutrient digestibility in hepu Geese[J].Chinese Journal of Animal Nutrition,2010,22(3):586-594.
[22]Matsumotoa H,Nogib T,Tabuchib I,et al.The SNPs in the promoter regionsof the bovine FADS2and FABP4genes are associated with beef quality traits[J].Livestock Science,2014,163:34-40.
[23]Gao Y,Zhang Y H,Zhang S,et al.Association of A-FABP gene polymorphism in intron 1with meat quality traits in Junmu No.1white swine[J].Gene,2011,11:170-173.
[24]Xu Q L,Tang G W,Zhang Q L,et al.The FABP4gene polymorphism is associated with meat tenderness in three Chinese native sheep breeds[J].Czech Journal of Animal Science,2011,56:1-6.
[25]Oh D Y,Lee Y S,La B M,et al.Identification of the SNP(Single Nucleotide Polymorphism)for fatty acid composition associated with beef flavorrelated FABP4(Fatty Acid Binding Protein 4)in Korean cattle[J].Animal Science,2012,25:913-920.
[26]Li M X,Sun X M,Hua L S,et al.SIRT1gene polymorphisms are associated with growth traits in Nanyang cattle[J].Molecular and Cellular Probes,2013,27:215-220.
[27]Cao F,Zhang H,Feng J,et al.Association study of three microsatellite polymorphisms located in introns 1,8,and 9 of DISC1 with schizophrenia in the Chinese Han population[J].Genet Test Mol Biomarkers,2013,17:407-411.
[2]Binas B,Danneberg H,McWhir J,et al.Requirement for the heart-type fatty acid binding protein incardiac fatty acid utilization[J].The FASEB Journal,2013,8:805-812.
[3]Caglar O,Hasim A,Cevat Y,et al.Heart-type fatty acid binding protein and cardiac-troponin I may have a diagnostic value in electrocution:A rat model[J].Journal of Forensic and Legal Medicine,2016,39:76-79.
[4]Chaap F G,Binas B,Danneberg H,et al.Impaired long-chain fatty acid utilization by cardiamyo-cytes isolated from mice lacking the heart-type fatty acid binding protein gene[J].Circulation research,1985,4:329-337.
[5]Kralisch S,Fasshauer M.Adipocyte fatty acid binding protein:a novel adipokine involved in the pathogenesis of metabolic and vascular disease?[J].Diabetologia,2013,1:10-21.
[6]Furuhashi M,Furuhashi M,Saitoh S,et al.Fatty acid-binding protein 4(FABP4):pathophysiological insights and potent clinical biomarker of metabolic and cardiovascular diseases[J].Clinical Medicine Insights:Cardiology,2015,3:23-33.
[7]Piegelman B M,Frank M,Green H.Molecular cloning of mRNA from 3T3adipocytes regulation of mRNA content for glycerop hosphate dehydrogenase and other differentiation-dependent proteins during adipocyte development[J].The Journal of Biologica Chemistry,1983,258(16):10083-10089.
[8]Haunerland N H,Spener F.Fatty acid-binding proteins-insights from genetic manipulations[J].Progress in Lipid Research,2004,43(4):328-349.
[9]Hotamisligil G S,Johnson R S,Distel R J,et al.Uncoupling of obesity from insulin resistance through a targeted mutation in aP2,the adipocyte fatty acidbinding protein[J].Science,1996,274(5291):1377-1379.
[10]Young C K,Yong K C,Won Y L,et al.Serum adipocyte-specific fatty acidbinding protein is associated with nonalcoholic fatty liver disease in apparently healthy subjects[J].The Journal of Nutritional Biochemistry,2011,22(3):289-292.
[11]Ysal K T,Scheja L,Wiesbrock S M,et al.Improved glucose and lipid metabolism in genetically obese mice lacking aP2[J].Endocrinology,2000,141(9):3388-3396.
[12]谈笑.FABP4对3T3-L1脂肪细胞中脂肪酸代谢和线粒体功能的影响[D].陕西杨凌:西北农林科技大学,2014.Tan X.Regulation of FABP4on fatty acid metabolism and mitochondrial function in 3T3-L1 adipocytes[D].Yangling,Shaanxi:Northwest A&F University,2014.
[13]桂林生,昝林森,王洪宝,等.SIRT1和SIRT2基因多态性及合并基因型与秦川牛肉用性状的关联分析[J].畜牧兽医学报,2015,46(10):1741-1749.Gui L S,Zan L S,Wang H B,et al.Effects of single and combined genotypes of SIRT1and SIRT2genes on meat quality traits in Qinchuan cattle[J].Chinese Journal of Animal and Veterinary Sciences,2015,46(10):1741-1749.
[14]Hardy H G.Mendelian proportions in a mixed population[J].Science,1908(28):49-50.
[15]Flint G S A.Structure of linkage disequilibrium in plants[J].Annu Rev Plant Biol,2003(54):357-374.
[16]李娟,杜新华,陈燕,等.牛FABP3转基因小鼠的遗传及表达特性研究[J].畜牧兽医学报,2015,46(10):1750-1758.Li J,Du X H,Chen Y,et al.Study on genetic characteristic and expression of bovine FABP3gene in transgenic mice[J].Chinese Journal of Animal and Veterinary Sciences,2015,46(10):1750-1758.
[17]Ye M H,Chen J L,Zhao G P,et al.Associations of A-FABP and H-FABP markers with the contnt of intramuscular fat in Beijing-You chicken[J].Anim Biotechnol,2010,21(1):14-24.
[18]Cho K H,Kim M J,Jeon G J,et al.Association of genetic variants for FABP3gene with back-fat thickness and intramuscular fat content in pig[J].Molecular Biology Reports,2011,38:2161-2166.
[19]Li X P,Kim S W,Choi J S,et al.Investigation of porcine FABP3and LEPR gene polymorphisms and mRNA expression for variation in intramuscular fat content[J].Molecular Biology Reports,2010,37:3931-3939.
[20]Blecha I M Z,Siqueira F,Ferreira A B R,et al.Identification and evaluation of polymorphisms in FABP3and FABP4in beef cattle[J].Genetics and Molecular Research,2015,14(4):16353-16363.
[21]Zhang J,Yang J H,Huang Y L,et al.Effects of different diet forms on growth performance and nutrient digestibility in hepu Geese[J].Chinese Journal of Animal Nutrition,2010,22(3):586-594.
[22]Matsumotoa H,Nogib T,Tabuchib I,et al.The SNPs in the promoter regionsof the bovine FADS2and FABP4genes are associated with beef quality traits[J].Livestock Science,2014,163:34-40.
[23]Gao Y,Zhang Y H,Zhang S,et al.Association of A-FABP gene polymorphism in intron 1with meat quality traits in Junmu No.1white swine[J].Gene,2011,11:170-173.
[24]Xu Q L,Tang G W,Zhang Q L,et al.The FABP4gene polymorphism is associated with meat tenderness in three Chinese native sheep breeds[J].Czech Journal of Animal Science,2011,56:1-6.
[25]Oh D Y,Lee Y S,La B M,et al.Identification of the SNP(Single Nucleotide Polymorphism)for fatty acid composition associated with beef flavorrelated FABP4(Fatty Acid Binding Protein 4)in Korean cattle[J].Animal Science,2012,25:913-920.
[26]Li M X,Sun X M,Hua L S,et al.SIRT1gene polymorphisms are associated with growth traits in Nanyang cattle[J].Molecular and Cellular Probes,2013,27:215-220.
[27]Cao F,Zhang H,Feng J,et al.Association study of three microsatellite polymorphisms located in introns 1,8,and 9 of DISC1 with schizophrenia in the Chinese Han population[J].Genet Test Mol Biomarkers,2013,17:407-411.