静原鸡ELOVL2基因遗传多态性研究
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摘要
为了探讨静原鸡极长链脂肪酸延伸酶蛋白2(elongation of very-long-chain fatty acids 2,ELOVL2)基因的遗传特性,确定核苷酸的变异位点,本研究采用PCR-SSCP及DNA测序技术对静原鸡ELOVL2基因进行多态性检测。结果显示,静原鸡ELOVL2基因外显子6无多态性,内含子2和6呈中度多态。在ELOVL2基因内含子2扩增片段上共检测到5种基因型:A_2A_2、B_2B_2、C2C2、D2D2和A_2C2,4种等位基因:A_2、B_2、C2和D2,其中A_2为优势等位基因(0.62),A_2A_2基因型为优势基因型(0.54);3个SNPs位点:g.63372228+4918G>A的转换、g.63372228+5024T>C的转换和g.63372228+4928C>A的颠换。在E2e6I6扩增片段上共检测到3种基因型:A_1A_1、B_1B_1和A_1B_1,2种等位基因:A_1和B_1,其中A_1为优势等位基因(0.67),A_1A_1基因型为优势基因型(0.54),等位基因B_1存在g.63388000+748G>C的颠换。群体遗传学分析发现,静原鸡ELOVL2基因内含子2、6遗传纯合度均较高,且偏离了Hardy-Weinberg平衡(P<0.05)。本研究结果表明,静原鸡ELOVL2基因内含子2和6序列突变较高,且呈中度多态,表现出纯合子优势,外显子6上无突变。
In order to study the genetic characteristics and nucleotide polymorphism sites of elongation of very-long-chain fatty acids 2(ELOVL2)gene in Jingyuan chicken,the polymorphisms of ELOVL2 gene were detected by polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP)and DNA sequence.The results showed that there was no polymorphism of ELOVL2 gene exon 6,there were moderate polymorphism of introns 2 and 6 in Jingyuan chicken.The total of 5 genotypes(A_2 A_2,B_2 B_2,C2 C2,D2 D2 and A_2 C2),4 alleles(A_2,B_2,C2 and D2)and3 SNPs loci were detected on the intron 2 amplified fragment of ELOVL2 gene.The 3 SNPs were g.63372228+4918 G>A transformation,g.63372228+5024 T>C transformation and g.63372228+4928 C>A transversion.A_2 was the dominant allele(0.62),and A_2 A_2 was the dominant genotype(0.54).The total of 3 genotypes(A_1 A_1,B_1 B_1 and A_1 B_1)and 2 alleles(A_1 and B_1)were detectedon E2 e6 I6 amplified fragments.A_1 was the dominant allele(0.67),A_1 A_1 was the dominant genotype(0.54),and only allele B_1 had a g.63388000+748 G>C transversion.Population genetics analysis showed that genetic homozygosity of ELOVL2 gene introns 2 and 6 in Jingyuan chicken were higher and deviation from Hardy-Weinberg equilibrium(P<0.05).The results showed that the mutations of the ELOVL2 gene introns 2 and 6 in the Jingyuan chicken were high,which were moderate polymorphism,showing the homozygous advantage and no mutation on the exon 6.
In order to study the genetic characteristics and nucleotide polymorphism sites of elongation of very-long-chain fatty acids 2(ELOVL2)gene in Jingyuan chicken,the polymorphisms of ELOVL2 gene were detected by polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP)and DNA sequence.The results showed that there was no polymorphism of ELOVL2 gene exon 6,there were moderate polymorphism of introns 2 and 6 in Jingyuan chicken.The total of 5 genotypes(A_2 A_2,B_2 B_2,C2 C2,D2 D2 and A_2 C2),4 alleles(A_2,B_2,C2 and D2)and3 SNPs loci were detected on the intron 2 amplified fragment of ELOVL2 gene.The 3 SNPs were g.63372228+4918 G>A transformation,g.63372228+5024 T>C transformation and g.63372228+4928 C>A transversion.A_2 was the dominant allele(0.62),and A_2 A_2 was the dominant genotype(0.54).The total of 3 genotypes(A_1 A_1,B_1 B_1 and A_1 B_1)and 2 alleles(A_1 and B_1)were detectedon E2 e6 I6 amplified fragments.A_1 was the dominant allele(0.67),A_1 A_1 was the dominant genotype(0.54),and only allele B_1 had a g.63388000+748 G>C transversion.Population genetics analysis showed that genetic homozygosity of ELOVL2 gene introns 2 and 6 in Jingyuan chicken were higher and deviation from Hardy-Weinberg equilibrium(P<0.05).The results showed that the mutations of the ELOVL2 gene introns 2 and 6 in the Jingyuan chicken were high,which were moderate polymorphism,showing the homozygous advantage and no mutation on the exon 6.
引文
[1]李维,祖盘玉,李洪林,等.赤水乌骨鸡肌肉生长抑制素基因多态性及其与肉质性状的关联分析[J].中国畜牧兽医,2017,44(4):1115-1121.LI W,ZU P Y,LI H L,et al.Polymorphisms of MSTN gene and its association with meat quality traits in Chishui Black-bone chicken[J].China Animal Husbandry&Veterinary Medicine,2017,44(4):1115-1121.(in Chinese)
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[12]BOTOLIN D,WANG Y,CHRISTIAN B,et al.Docosahexaneoic acid(22∶6,n-3)regulates rat hepatocyte SREBP-1nuclear abundance by Erk and 26Sproteasome-dependent pathways[J].Journal of Lipid Research,2006,47(1):181-92.
[13]李响,丁珍,吴义霞,等.ELOVL2基因多态性与乳母乳汁DHA水平的关联性分析[J].吉林大学学报(医学版),2015,41(6):1230-1234.LI X,DING Z,WU Y X,et al.Association analysis on polymorphisms of ELOVL2gene and DHA level in breast milk of lactating mothers[J].Journal of Jilin University(Medical Edition),2015,41(6):1230-1234.(in Chinese)
[14]郭鹏程,戴珊珊,杨浩然,等.ELOVL5基因遗传变异与中国荷斯坦奶牛乳质性状的关联分析及功能验证[J].中国兽医学报,2017,37(4):741-745.GUO P C,DAI S S,YANG H R,et al.elovl5gene variants reveal significant effects of milk quality in holstein cows[J].Chinese Journal of Veterinary Science,2017(4):741-745.(in Chinese)
[15]TANAKA T,SHEN J,ABECASIS G R,et al.Genome-wide association study of plasma polyunsaturated fatty acids in the InCHIANTI study[J].PLoS Genetics,2009,5(1):e1000338.
[16]ILLIG T,GIEGER C,ZHAI G,et al.A genome-wide perspective of genetic variation in human metabolism[J].Nature Genetics,2010,42(2):137-141.
[17]LEMAITRE R N,TANAKA T,TANG W,et al.Genetic loci associated with plasma phospholipid n-3fatty acids:A meta-analysis of genome-wide association studies from the CHARGE consortium[J].PLoS Genetics,2011,7(7):e1002193.
[18]邵丹,王来娣,张蕊,等.鹅肥肝形成相关基因的研究进展[J].中国畜牧兽医,2012,39(9):42-46.SHAO D,WANG L D,ZHANG R,et al.Research progress of genes related to fatty liver formation in goose[J].China Animal Husbandry&Veterinary Medicine,2012,39(9):42-46.(in Chinese)
[19]孙晓艳.小鼠elovl2与△6desaturase及线虫sfat1三基因共转促进EPA向DHA转化的研究[D].西安:陕西师范大学,2012.SUN X Y.Co transfection of mouse elovl2and△6desaturase and sfat1of nematode promote the transformation of EPA to DHA[D].Xi’an:Shaanxi Normal University,2012.(in Chinese)
[20]母童,张娟,赵平,等.静原鸡ELOVL2和ELOVL5基因表达的组织特异性研究[J].浙江农业学报,2017,29(8):1290-1296.MU T,ZHANG J,ZHAO P,et al.Tissue-specific expression analysis of ELOVL2and ELOVL5genes in Jingyuan chicken[J].Acta Agriculturae Zhejiangensis,2017,29(8):1290-1296.(in Chinese)
[21]夏德佳,孙琳,史杰萍,等.多不饱和脂肪酸代谢限速酶基因多态性与吉林省汉族偏执型精神分裂症的病例-对照研究[J].卫生研究,2011,40(2):154-157.XIA D J,SUN L,SHI J P,et al.A case-control study between gene polymorphisms of polyunsaturated fatty acids metabolic rate-limiting enzymes and paranoid schizophrenia of Han ethnicity in Jilin province[J].Journal of Hygiene Research,2011,40(2):154-157.(in Chinese)
[22]邢华苑,王颜刚.ELOVL6基因rs9997926位点多态性与2型糖尿病病人胰岛素抵抗的相关性[J].青岛大学医学院学报,2017,53(1):84-86.XING H Y,WANG Y G.Correlation between ELOVL6gene rs9997926polymorphism and insulin resistance in type 2diabetes[J].Acta Academiae Medicinae Qingdao Universitatis,2017,53(1):84-86.(in Chinese)
[23]尉阳,王蕾蕾,金镇.ELOVL6基因单核苷酸多态性与妊娠期糖尿病的相关性[J].解剖科学进展,2015,21(2):171-173.WEI Y,WANG L L,JIN Z.Association between single nucleotide polymorphism of ELOVL6and gestational diabetes mellitus[J].Progress of Anatomical Sciences,2015,21(2):171-173.(in Chinese)
[24]LI X,GAN Z W,DING Z,et al.Genetic variants in the ELOVL5but not ELOVL2gene associated with polyunsaturated fatty acids in Han Chinese breast milk[J].Biomedical and Environmental Sciences,2017,30(1):64-67.
[2]顾亚玲,张娟,何生虎.静原鸡品种选育与饲养管理[M].宁夏:阳光出版社,2017.GU Y L,ZHANG J,HE S H.Breed Selection and Feeding Management in Jingyuan Chicken[M].Ningxia:Sunshine Press,2017.(in Chinese)
[3]ZHANG J,ZHAO S,LEI Z,et al.Identification of polymorphisms in the bovine paraoxonase 1gene[J].Journal of Animal&Veterinary Advances,2013,12(2):229-233.
[4]NAGANUMA T,SATO Y,SASSA T,et al.Biochemical characterization of the very long-chain fatty acid elongase ELOVL7[J].FEBS Letters,2011,585(20):3337-3341.
[5]LEONARD A E,PEREIRA S L,SPRECHER H,et al.Elongation of long-chain fatty acids[J].Progress in Lipid Research,2004,43(1):36.
[6]JAKOBSSON A,WESTERBERG R,JACOBSSON A.Fatty acid elongases in mammals:Their regulation and roles in metabolism[J].Progress in Lipid Research,2006,45(3):237-249.
[7]GUILLOU H,ZADRAVEC D,MARTIN P G,et al.The key roles of elongases and desaturases in mammalian fatty acid metabolism:Insights from transgenic mice[J].Progress in Lipid Research,2010,49(2):186-199.
[8]SCAR MONROIG,ROTLLANT J,CERD-REVERTER J M,et al.Expression and role of Elovl4elongases in biosynthesis of very long-chain fatty acids during zebrafish Danio rerio,early embryonic development[J].Biochimica et Biophysica Acta,2010,1801(10):1145.
[9]LEONARD A E,KELDER B,BOBIK E G,et al.Identification and expression of mammalian long-chain PUFA elongation enzymes[J].Lipids,2002,37(8):733-740.
[10]KOBAYASHI T,ZADRAVEC D,JACOBSSON A.ELOVL2 overexpression enhances triacylglycerol synthesis in 3T3-L1and F442A cells[J].Febs Letters,2007,581(17):3157.
[11]WANG Y,BOTOLIN D,CHRISTIAN B,et al.Tissue-specific,nutritional,and developmental regulation of rat fatty acid elongases[J].Journal of Lipid Research,2005,46(4):706-715.
[12]BOTOLIN D,WANG Y,CHRISTIAN B,et al.Docosahexaneoic acid(22∶6,n-3)regulates rat hepatocyte SREBP-1nuclear abundance by Erk and 26Sproteasome-dependent pathways[J].Journal of Lipid Research,2006,47(1):181-92.
[13]李响,丁珍,吴义霞,等.ELOVL2基因多态性与乳母乳汁DHA水平的关联性分析[J].吉林大学学报(医学版),2015,41(6):1230-1234.LI X,DING Z,WU Y X,et al.Association analysis on polymorphisms of ELOVL2gene and DHA level in breast milk of lactating mothers[J].Journal of Jilin University(Medical Edition),2015,41(6):1230-1234.(in Chinese)
[14]郭鹏程,戴珊珊,杨浩然,等.ELOVL5基因遗传变异与中国荷斯坦奶牛乳质性状的关联分析及功能验证[J].中国兽医学报,2017,37(4):741-745.GUO P C,DAI S S,YANG H R,et al.elovl5gene variants reveal significant effects of milk quality in holstein cows[J].Chinese Journal of Veterinary Science,2017(4):741-745.(in Chinese)
[15]TANAKA T,SHEN J,ABECASIS G R,et al.Genome-wide association study of plasma polyunsaturated fatty acids in the InCHIANTI study[J].PLoS Genetics,2009,5(1):e1000338.
[16]ILLIG T,GIEGER C,ZHAI G,et al.A genome-wide perspective of genetic variation in human metabolism[J].Nature Genetics,2010,42(2):137-141.
[17]LEMAITRE R N,TANAKA T,TANG W,et al.Genetic loci associated with plasma phospholipid n-3fatty acids:A meta-analysis of genome-wide association studies from the CHARGE consortium[J].PLoS Genetics,2011,7(7):e1002193.
[18]邵丹,王来娣,张蕊,等.鹅肥肝形成相关基因的研究进展[J].中国畜牧兽医,2012,39(9):42-46.SHAO D,WANG L D,ZHANG R,et al.Research progress of genes related to fatty liver formation in goose[J].China Animal Husbandry&Veterinary Medicine,2012,39(9):42-46.(in Chinese)
[19]孙晓艳.小鼠elovl2与△6desaturase及线虫sfat1三基因共转促进EPA向DHA转化的研究[D].西安:陕西师范大学,2012.SUN X Y.Co transfection of mouse elovl2and△6desaturase and sfat1of nematode promote the transformation of EPA to DHA[D].Xi’an:Shaanxi Normal University,2012.(in Chinese)
[20]母童,张娟,赵平,等.静原鸡ELOVL2和ELOVL5基因表达的组织特异性研究[J].浙江农业学报,2017,29(8):1290-1296.MU T,ZHANG J,ZHAO P,et al.Tissue-specific expression analysis of ELOVL2and ELOVL5genes in Jingyuan chicken[J].Acta Agriculturae Zhejiangensis,2017,29(8):1290-1296.(in Chinese)
[21]夏德佳,孙琳,史杰萍,等.多不饱和脂肪酸代谢限速酶基因多态性与吉林省汉族偏执型精神分裂症的病例-对照研究[J].卫生研究,2011,40(2):154-157.XIA D J,SUN L,SHI J P,et al.A case-control study between gene polymorphisms of polyunsaturated fatty acids metabolic rate-limiting enzymes and paranoid schizophrenia of Han ethnicity in Jilin province[J].Journal of Hygiene Research,2011,40(2):154-157.(in Chinese)
[22]邢华苑,王颜刚.ELOVL6基因rs9997926位点多态性与2型糖尿病病人胰岛素抵抗的相关性[J].青岛大学医学院学报,2017,53(1):84-86.XING H Y,WANG Y G.Correlation between ELOVL6gene rs9997926polymorphism and insulin resistance in type 2diabetes[J].Acta Academiae Medicinae Qingdao Universitatis,2017,53(1):84-86.(in Chinese)
[23]尉阳,王蕾蕾,金镇.ELOVL6基因单核苷酸多态性与妊娠期糖尿病的相关性[J].解剖科学进展,2015,21(2):171-173.WEI Y,WANG L L,JIN Z.Association between single nucleotide polymorphism of ELOVL6and gestational diabetes mellitus[J].Progress of Anatomical Sciences,2015,21(2):171-173.(in Chinese)
[24]LI X,GAN Z W,DING Z,et al.Genetic variants in the ELOVL5but not ELOVL2gene associated with polyunsaturated fatty acids in Han Chinese breast milk[J].Biomedical and Environmental Sciences,2017,30(1):64-67.