静原鸡ELOVL5基因遗传多样性研究
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摘要
为确定静原鸡ELOVL5基因遗传变异位点,探讨其遗传特性,本研究利用单链构象多态性分析(PCR-SSCP)及DNA测序技术对静原鸡ELOVL5基因进行多态性检测。结果表明,静原鸡ELOVL5基因exon2和ex-on5均无多态性;在intron2扩增片段上共检测到1个SNP位点(g. 88620433+1683G> A),其表现为2种基因型(AA、AG)。其中AA为优势基因型,基因型频率为0. 84,A为优势等位基因,基因频率为0. 92。在248个静原鸡群体中未发现纯合GG型个体。群体遗传学分析表明,该位点纯合度较高(0. 84),PIC为0. 14(PIC <0. 25),呈低度多态。卡方适合性检验结果表明,静原鸡ELOVL5基因intron2突变未偏离Hardy-Weinberg平衡(P> 0. 05)。综上,静原鸡ELOVL5基因intron2多态性较低,exon2和exon5未发生变异。
Nucleotide polymorphism and site genetic characteristics of ELOVL5 gene in Jingyuan chicken were studied. Polymorphisms of ELOVL5 gene were detected by polymerase chain reaction-single strand conformation polymorphism( PCR-SSCP) and DNA sequence. The results showed that the ELOVL5 gene exon2 and exon5 had no polymorphism. The total of 1 SNP( g. 88620433 + 1683 G > A),2 genotypes were detected on the intron2 amplified fragment of ELOVL5 gene( AA,AG),genotype AA and allele A was dominant,the frequency was 0. 84 and 0. 92. No homozygous GG type individual was found in 248 populations of Jingyuan chicken. Population genetic analysis showed that the homozygosity of the locus was higher( 0. 84) and PIC was 0. 14( PIC < 0. 25),showing low polymorphism. The results showed that the intron2 mutation of ELOVL5 gene in Jingyuan chicken did not deviate from Hardy-Weinberg equilibrium by χ2 test( P > 0. 05). To sum up,the amplified fragment of the intron2 gene of Jingyuan chicken had low polymorphism,and there was no mutation in exon2 and exon5.
Nucleotide polymorphism and site genetic characteristics of ELOVL5 gene in Jingyuan chicken were studied. Polymorphisms of ELOVL5 gene were detected by polymerase chain reaction-single strand conformation polymorphism( PCR-SSCP) and DNA sequence. The results showed that the ELOVL5 gene exon2 and exon5 had no polymorphism. The total of 1 SNP( g. 88620433 + 1683 G > A),2 genotypes were detected on the intron2 amplified fragment of ELOVL5 gene( AA,AG),genotype AA and allele A was dominant,the frequency was 0. 84 and 0. 92. No homozygous GG type individual was found in 248 populations of Jingyuan chicken. Population genetic analysis showed that the homozygosity of the locus was higher( 0. 84) and PIC was 0. 14( PIC < 0. 25),showing low polymorphism. The results showed that the intron2 mutation of ELOVL5 gene in Jingyuan chicken did not deviate from Hardy-Weinberg equilibrium by χ2 test( P > 0. 05). To sum up,the amplified fragment of the intron2 gene of Jingyuan chicken had low polymorphism,and there was no mutation in exon2 and exon5.
引文
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[18]覃川杰,文正勇,袁登越,等.瓦氏黄颡鱼(Pelteobagrus vachellii)脂肪酸去饱和酶FAD2和延伸酶ELOVL5的克隆及表达分析[J].海洋与湖沼,2017,48(4):884-893.QIN C J,WEN Z Y,YUAN D Y,et al.Molecular cloning and expression of fatty acid desaturase and elongase genes in darkbarbel catfish Pelteobagrus vachellii[J].Oceanologia Et Limnologia Sinica,2017,48(4):884-893.(in Chinese with English abstract)
[19]谭小音,金丽,谢德琼,等.八眉猪SLA-DRA基因第4外显子多态性分析[J].生物技术,2015,25(5):463-467.TAN X Y,JIN L,XIE D Q,et al.Polymorphism of SLA-DRA exon 4 in Bamei pigs[J].Biotechnology,2015,25(5):463-467.(in Chinese with English abstract)
[20]REN H R,YU J,XU P,et al.Single nucleotide polymorphisms of6-desaturase and ELOVL5 segments and their associations with common carp(Cyprinus carpio)growth traits[J].Genetics&Molecular Research,2015,14(4):12848-12854.
[21]SHI H B,DU Y,ZHANG C H,et al.Fatty acid elongase 5(ELOVL5)alters the synthesis of long-chain unsaturated fatty acids in goat mammary epithelial cells[J].Journal of Dairy Science,2018,101(5):4586-4587.
[22]ZHANG M,LI C C,LI F,et al.Estrogen promotes hepatic synthesis of long-chain polyunsaturated fatty acids by regulating ELOVL5 at post-transcriptional level in laying hens[J].International Journal of Molecular Sciences,2017,18(7):pii:E1405.
[2]姜琳琳,刘华贵,齐德生,等.鸡肉中脂肪酸的研究进展[J].肉类研究,2006(1):37-40.JIANG L L,LIU H G,QI D S,et al.Review on fatty acid in muscle tissues of chicken[J].Meat Research,2006(1):37-40.(in Chinese with English abstract)
[3]母童,顾亚玲,赵平,等.静原鸡ELOVL2基因遗传多态性研究[J].中国畜牧兽医,2018,45(3):730-737.MU T,GU Y L,ZHAO P,et al.Study on polymorphism of ELOVL2 gene in Jingyuan chicken[J].China Animal Husbandry&Veterinary Medicine,2018,45(3):730-737.(in Chinese with English abstract)
[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]唐慧,潘志雄,卢立志,等.超长链脂肪酸延伸酶家族的功能及表达调控[J].生命的化学,2009,29(6):898-901.TANG H,PAN Z X,LU L Z,et al.The function and expression regulation of elongases of very long chain fatty acids[J].Chemistry of Life,2009,29(6):898-901.(in Chinese with English abstract)
[6]SHIKAMA A,SHINOZAKI H,TAKEUCHI Y,et al.Identification of human ELOVL5 enhancer regions controlled by SREBP[J].Biochemical&Biophysical Research Communications,2015,465(4):857-863.
[7]QIN Y,DALEN K T,GUSTAFSSON J,et al.Regulation of hepatic fatty acid elongase 5 by LXRα-SREBP-1c[J].Biochimica et Biophysica Acta,2009,1791(2):140-147.
[8]WANG Y,TORRESGONZALEZ M,TRIPATHY S,et al.Elevated hepatic fatty acid elongase-5 activity affects multiple pathways controlling hepatic lipid and carbohydrate composition[J].Journal of Lipid Research,2008,49(7):1538-1552.
[9]王海燕,苏玉虹.编码极长链脂肪酸延长酶基因家族的结构及其产物的功能[J].生命的化学,2005,25(1):29-31.WANG H Y,SU Y H.The structure and function of the coding long chain fatty acid gene family[J].Chemistry of Life,2005,25(1):29-30.(in Chinese)
[10]邵亚明,罗发兴,田泽维,等.慢性乙型肝炎基因治疗若干进展[J].生命的化学,2005,25(1):31-33.SHAO Y M,LUO F X,TIAN Z W,et al.Advances in gene therapy of chronic hepatitis B[J].Chemistry of Life,2005,25(1):31-33.(in Chinese)
[11]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 ELOVL2 and ELOVL5 genes in Jingyuan chicken[J].Acta Agriculturae Zhejiangensis,2017,29(8):1290-1296.(in Chinese with English abstract)
[12]MATSUMOTO H,SHIMIZU Y,TANAKA A,et al.The SNP in the promoter region of the bovine ELOVL5 gene influences economic traits including subcutaneous fat thickness[J].Molecular Biology Reports,2013,40(4):3231-3237.
[13]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.ELOVL5 gene variants reveal significant effects of milk quality in Holstein cows[J].Chinese Journal of Veterinary Science,2017,37(4):741-745.(in Chinese with English abstract)
[14]XIANG L I,GAN Z W,DING Z,et al.Genetic variants in the ELOVL5 but not ELOVL2 gene associated with polyunsaturated fatty acids in han chinese breast milk[J].Biomedical and Environmental Sciences,2017,30(1):64-67.
[15]ZUO R,MAI K,XU W,et al.Molecular cloning,tissue distribution and nutritional regulation of a fatty acyl ELOVL5-like elongase in large yellow croaker,Larimichthys crocea[J].Aquaculture Research,2016,47(8):2393-2406.
[16]CARMONA-ANTONANZAS G,ZHENG X,TOCHER D R,et al.Regulatory divergence of homeologous atlantic salmon ELOVL5 genes following the salmonid-specific whole-genome duplication[J].Gene,2016,591(1):34-42.
[17]吴景,郑先虎,匡友谊,等.镜鲤脂肪酸延长酶5基因的克隆和表达分析[J].中国水产科学,2015,22(1):9-16.WU J,ZHENG X H,KUANG Y Y,et al.Cloning and characteristic analysis of fatty acid elongase 5 in mirror carp(Cyprinus carpio)[J].Journal of Fishery Sciences of China,2015,22(1):9-16.(in Chinese with English abstract)
[18]覃川杰,文正勇,袁登越,等.瓦氏黄颡鱼(Pelteobagrus vachellii)脂肪酸去饱和酶FAD2和延伸酶ELOVL5的克隆及表达分析[J].海洋与湖沼,2017,48(4):884-893.QIN C J,WEN Z Y,YUAN D Y,et al.Molecular cloning and expression of fatty acid desaturase and elongase genes in darkbarbel catfish Pelteobagrus vachellii[J].Oceanologia Et Limnologia Sinica,2017,48(4):884-893.(in Chinese with English abstract)
[19]谭小音,金丽,谢德琼,等.八眉猪SLA-DRA基因第4外显子多态性分析[J].生物技术,2015,25(5):463-467.TAN X Y,JIN L,XIE D Q,et al.Polymorphism of SLA-DRA exon 4 in Bamei pigs[J].Biotechnology,2015,25(5):463-467.(in Chinese with English abstract)
[20]REN H R,YU J,XU P,et al.Single nucleotide polymorphisms of6-desaturase and ELOVL5 segments and their associations with common carp(Cyprinus carpio)growth traits[J].Genetics&Molecular Research,2015,14(4):12848-12854.
[21]SHI H B,DU Y,ZHANG C H,et al.Fatty acid elongase 5(ELOVL5)alters the synthesis of long-chain unsaturated fatty acids in goat mammary epithelial cells[J].Journal of Dairy Science,2018,101(5):4586-4587.
[22]ZHANG M,LI C C,LI F,et al.Estrogen promotes hepatic synthesis of long-chain polyunsaturated fatty acids by regulating ELOVL5 at post-transcriptional level in laying hens[J].International Journal of Molecular Sciences,2017,18(7):pii:E1405.