山羊Wnt5A基因序列变异及其组织表达特征  

Sequence Variation and Tissue Expression Characteristics of Wnt5A Gene in Goat(Capra hircus)

作　　者：李少斌 张瑞国 何兆华 赵鹏飞 赵芳芳 王继卿 刘秀 胡江 罗玉柱 LI Shao-Bin;ZHANG Rui-Guo;HE Zhao-Hua;ZHAO Peng-Fei;ZHAO Fang-Fang;WANG Ji-Qing;LIU Xiu;HU Jiang;LUO Yu-Zhu(College of Animal Science and Technology/Gansu Key Laboratory of Herbivorous Animal Biotechnology,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物科学技术学院/甘肃省草食动物生物技术重点实验室,兰州730070

出　　处：《农业生物技术学报》2022年第7期1321-1328,共8页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(32060140);甘肃省杰出青年基金(21JR7RA857);甘肃农业大学伏羲青年英才项目(Gaufx-03Y04);甘肃省草食动物生物技术重点实验室开放课题(GKLAB-201702)。

摘　　要：羊绒的质量和产量决定着绒山羊(Capra hircus)的经济价值,而毛囊是调控羊绒生长和性状的直接组织,通过调控毛囊发育可实现羊绒性状的改良。无翅型鼠乳腺肿瘤病毒整合位点家族成员5A(wingless-type mouse mammary tumor virus integration site family member 5A,Wnt5A)基因是毛囊发育信号通路中的关键因子,在毛囊的发育过程中起着关键作用,但其在山羊中的研究还非常有限。为全面掌握山羊Wnt5A的分子遗传特征,本研究利用PCR-单链构象多态性(PCR-single strand conformation polymorphism,PCR-SSCP)结合测序,检测分析该基因在中卫山羊、柴达木绒山羊和陇东绒山羊群体中的变异特征;利用qPCR技术,分析Wnt5A在辽宁绒山羊和陇东绒山羊组织表达特征。结果显示:在陇东绒山羊、柴达木绒山羊和中卫山羊群体中,Wnt5A的exon 4扩增区存在2个突变位点,分别为c.334C>A和c.213A>G,形成A、B和C 3个等位基因并表现为AA、AB、AC、BB和BC 5种基因型。A和AB分别为柴达木绒山羊和陇东绒山羊中的优势等位基因和优势基因型,而中卫山羊中分别为B和BB。在该基因exon 6扩增区检测到由D和E 2个等位基因构成的DD、DE和EE 3种基因型,序列比对发现c.685-5C>T和c.685-55G>A 2个SNPs。3个群体中E均为优势等位基因,DE均为优势基因型。2个扩增区构成H1~H66种单倍型,其中频率最高的为H2。Wnt5A基因在辽宁绒山羊和陇东绒山羊的7种组织中均表达,品种间具有差异性,同时在皮肤中的表达差异显著,推测是由于Wnt5A基因的表达与羊绒性状相关造成的。该研究通过比较不同品种山羊Wnt5A基因的变异和表达特征,分析了该基因的分子遗传特征,为丰富山羊基因组内容及该基因的开发利用提供了基础资料。The quality and yield of cashmere determine the economic value of cashmere goats(Capra hircus),and hair follicle is the direct tissue to regulate the growth and properties of cashmere.The properties of cashmere can be improved by regulating the development of hair follicle.Wingless-type mouse mammary tumor virus integration site family,member 5A(Wnt5A)gene is an key member in the signaling pathway relates with hair follicle development and play a key role in hair follicle development,but its research in goat is still very limited.In order to well know the molecular genetic characteristics of goat Wnt5A,the variation of this gene was detected by combining PCR-single strand conformation polymorphism(PCR-SSCP)and sequencing in Zhongwei goats,Chaidamu Cashmere goats and Longdong Cashmere goats.The tissue expression characteristics of Wnt5a in Liaoning Cashmere goats and Longdong Cashmere goats were analyzed by qPCR.The results showed that,in the 3 populations,there were 2 mutation sites in the Wnt5A exon 4amplification region,which were c.334c>A and c.213a>G,respectively.Three alleles(A,B and C)were formed and showed five genotypes(AA,AB,AC,BB and BC).In Chaidamu cashmere goat and Longdong cashmere goat,A was the dominant allele,AB was the dominant genotype,while B in Zhongwei goat was the dominant allele and BB was the dominant genotype.Two mutation sites c.685-5 C>T and c.685-55 G>A were detected in Wnt5A exon 6,forming 2 alleles D and E,forming 3 genotypes DD,DE and EE.E was the dominant allele and DE was the dominant genotype in the 3 populations.Six haplotypes of H1~H6 were constituted by 2 amplified regions,and the most common one was H2.Wnt5A gene was expressed in all these7 tissues of Liaoning Cashmere goat and Longdong Cashmere goat,with differences between breeds.At the same time,the expression difference in skin was significant,which speculated that resulted from the correlation between Wnt5A gene expression and the cashmere traits.By comparing the variation and expression characteristics of Wnt5A gene in

关 键 词：WNT5A 绒山羊 序列变异 特异性表达 

分 类 号：S826[农业科学—畜牧学]