大白猪MC4R、CDC16基因多态性及其与生长性状的关联分析
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摘要
本研究旨在利用高分辨率熔解曲线(high resolution melt,HRM)分型方法检测大白猪黑素皮质素受体(melanocortin-4 receptor,MC4R)和细胞分裂周期蛋白16(cell division cycle 16,CDC16)基因的多态性,并对其与大白猪的生长性状进行关联性分析。试验共采集246头大白猪的耳组织样提取基因组DNA,对小群体样本进行PCR扩增后,通过测序方法寻找两个基因在该群体中的多态性位点,针对已发现的多态性位点设计HRM专用引物,利用HRM方法对大群体进行多态性检测,将所得结果与大白猪生长性状进行关联性分析。结果表明,在MC4R基因序列中检测到1个突变位点2207A>G,2个等位基因:A和G,3个基因型:AA、AG和GG;在CDC16基因第18外显子上检测到了1个突变位点837T>C,2个等位基因:T和C,2种基因型:TT和CT。χ~2适合性检验结果显示,2个基因的基因型频率和等位基因频率在2个种群中分布差异不显著,符合Hardy-Weinberg平衡定律(P>0.05)。多态信息含量(PIC)分析显示,MC4R基因在2个群体中均处于中度多态(0.25 This study was aimed to detect the polymorphism of melanocortin-4 receptor(MC4 R) and cell division cycle protein 16(CDC16) in Large White pigs by HRM typing,and analyze its association with growth traits in Large White pigs.A total of 246 Large White pigs ear tissues were collected as research objects,DNA was extracted using the AxyPrep Genomic DNA Extraction Kit.After PCR amplification of small population samples,the polymorphic positions of MC4R and CDC16 genes in the population were searched by sequencing.According to the findings,the polymorphic sites were designed for HRM-specific primers.The HRM method was used to detect the polymorphism in large populations,and the results were correlated with growth traits in Large White pigs.The results showed that one mutation site 2207 A>G,two alleles(A and G) and three genotypes(AA,AG and GG) were detected in the MC4R gene sequence.There was a mutation site 837 T>C,two alleles(T and C),two genotypes(TT and CT) in the 18 th exon of CDC16 gene.The χ~2 test results showed that the genotype frequency and gene frequency of MC4R and CDC16 genes were not significant difference between the two populations,which was consistent with the Hardy-Weinberg equilibrium(P>0.05).The polymorphic information content(PIC) result showed that MC4R gene was moderate polymorphic(0.25
引文
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[18] CHEN F,LIU N F.A 10-gene expression signature of Notch pathway predicts recurrencein ovarian carcinoma[J].Oncology Letters,2015,10(3):1704-1708.
[19] JEREMIAN R,CHEN Y A,LUCA V D,et al.Investigation of correlations between DNA methylation,suicidal behavior and aging[J].Bipolar Disorders,2017,19(1):32-40.
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[2] 闫德超.大白猪MC4R、IGF2和VRTN基因的多态性及其与生产性状的关联分析[D].武汉:华中农业大学,2017. YAN D C.Polymorphism and association analysis of MC4R,IGF2 and VRTN genes with production traits in Large White pigs[D].Wuhan:Huazhong Agricultural University,2017.(in Chinese)
[3] 马海明.猪4个新基因的分离、物理定位及MyoG基因SNP研究[D].长沙:湖南农业大学,2005. MA H M.Four new genes’ isolation,physical localization and MyoG gene SNP study of pigs[D].Changsha:Hunan Agricultural University,2005.(in Chinese)
[4] 杜海波.猪CDC16、EFNB2基因的多态性及其与部分经济性状相关性研究[D].长沙:湖南农业大学,2011. DU H B.Studies on CDC16 gene,EFNB2 gene polymorphisms and relations with partial economic traits in pigs[D].Changsha:Hunan Agricultural University,2005.(in Chinese)
[5] 杨公社.猪生产学[M].第14版.北京:中国农业出版社,2013. YANG G S.Swine Science[M].14th Edition.Beijing:China Agriculture Press,2013.(in Chinese)
[6] 嘎利兵嘎.高分辨率熔解曲线(HRM)技术在动物病毒基因分型中的研究及应用[D].北京:中国农业大学,2015. GA L B G.Research and application of high resolution melting curve analysis in the genotyping and detection of animal viruses[D].Beijing:China Agricultural University,2015.(in Chinese)
[7] 陈焱森,沈永巧,祁梦凡,等.应用高分辨溶解曲线(HRM)技术分析猪微卫星多态性[J].畜牧与兽医,2018,50(1):9-14. CHEN Y S,SHEN Y Q,QI M F,et al.Application of high-resolution melting curve analysis on polymorphism of SSR markers in pigs[J].Animal Husbandry and Veterinary Medicine,2018,50(1):9-14.(in Chinese)
[8] 易恒洁,李辉,赵忠海,等.高坡猪肌肉生长抑制素基因多态性及其与肉质性状的相关性分析[J].中国畜牧兽医,2017,44(4):1102-1107. YI H J,LI H,ZHAO Z H,et al.The polymorphisms of MSTN gene and its relationship with meat quanlity traits in Gaopo pig[J].China Animal Husbandry & Veterinary Medicine,2017,44(4):1102-1107.(in Chinese)
[9] 徐炜琳.松辽黑猪四种基因多态性与背膘厚的关联分析[D].长春:吉林农业大学,2017. XU W L.The association of polymorphisms of four genes with the backfat in Songliao Black swine[D].Changchun:Jilin Agricultural University,2017.(in Chinese)
[10] 韩雪蕾,乔瑞敏,李素雅,等.MC4R基因遗传变异与猪生长性状的相关性分析[J].猪业科学,2015,32(11):110-112. HAN X L,QIAO R M,LI S Y,et al.Correlation analysis between genetic variation of MC4R gene and growth traits in pigs[J].Swine Industry Science,2015,32(11):110-112.(in Chinese)
[11] 黄李勇,赵雄,曼则热·朱尔丁,等.巴什拜羊MC4R基因多态性及其与生长性状的关联分析[J].草食家畜,2018,3:17-23.(in Chinese) HUANG L Y,ZHAO X,MAN Z R·Z E D,et al.Association analysis of single nucleotide polymormhism of MC4R gene with growth traits in Bashibai sheep[J].Grass-Feeding Livestock,2018,3:17-23.(in Chinese)
[12] 李天科.牦牛GDF-10、IHH、MC4R基因的多态与生长性状的相关性研究[D].兰州:甘肃农业大学,2014. LI T K.Study on the genetic polymorphism of the yak GDF-10,IHH and MC4R gene and its association with growth trait[D].Lanzhou:Gansu Agricultural University,2014.(in Chinese)
[13] 潘宝丽.貉子MC4R、POU1F1基因多态性及其与生长性状关系的研究[D].长春:吉林农业大学,2011. PAN B L.Polymorphisms of MC4R and POU1F1 genes and its associations with several growth traits in Raccoon dog[D].Changchun:Jilin Agricultural University,2011.(in Chinese)
[14] CHOI J S,JIN S K,JEONG Y H,et al.Relationships between single nucleotide polymorphism markers and meat quality traits of Duroc breeding stocks in Korea[J].Asian-Australasian Journal of Animal Sciences,2016,29(9):1229-1238.
[15] HIROSE K,ITO T,FUKAWA K,et al.Evaluation of effects of multiple candidate genes (LEP,LEPR,MC4R,PIK3C3,and VRTN) on production traits in Duroc pigs[J].Animal Science Journal,2014,85(3):198-206.
[16] SWITONSKI M,MANKOWSKA M,SALAMON S.Family ofmelanocortin receptor (MCR) genes in mammals—mutations,polymorphisms and phenotypic effects[J].Journal of Applied Genetics,2013,54(4):461-472.
[17] FONTANESI L,BUTTAZZONI L,GALIMBERTI G,et al.Association between melanocortin 4 receptor (MC4R) gene haplotypes and carcass and production traits in Italian Large White pigs evaluated with a selective genotyping approach[J].Livestock Science,2013,157(1):48-56.
[18] CHEN F,LIU N F.A 10-gene expression signature of Notch pathway predicts recurrencein ovarian carcinoma[J].Oncology Letters,2015,10(3):1704-1708.
[19] JEREMIAN R,CHEN Y A,LUCA V D,et al.Investigation of correlations between DNA methylation,suicidal behavior and aging[J].Bipolar Disorders,2017,19(1):32-40.
[20] CHEN Q L,XIE B X,ZHU S S,et al.A TBC1D1 Ser231Ala-knockin mutation partially impairs AICAR- but not exercise-induced muscle glucose uptake in mice[J].Diabetologia,2017,60(2):336-345.
[21] FENTZ J,BIRK J B,JORDY A B,et al.AMPKα is critical for enhancing skeletal muscle fatty acid utilization during in vivo exercise in mice[J].FASEB Journal,2015,29(5):1725-1738.
[22] CHEN L,CHEN Q L,XIE B X,et al.Disruption of the AMPK-TBC1D1 nexus increases lipogenic gene expression and causes obesity in mice via promoting IGF1 secretion[J].Proceedings of the National Academy of Sciences of the United States of America,2016,113(26):7219-7224.