酉州乌羊皮肤组织ASIP基因的RACE克隆及分析  

RACE Cloning and Analysis of ASIP in Youzhou Dark Goat Skin Tissues

作　　者：付琳 孙晓燕[1,2] 任航行 李杰[1,2] 蒋婧 王高富[1,2] FU Lin;SUN Xiao-yan;REN Hang-xing;LI Jie;JIANG Jing;WANG Gao-fu(Chongqing Academy of Animal Sciences,Chongqing Rongchang 402460,China;Chongqing Engineering Research Center for Goat,Chongqing Rongchang 402460,China)

机构地区：[1]重庆市畜牧科学院,重庆荣昌402460 [2]重庆市山羊工程技术研究中心,重庆荣昌402460

出　　处：《西南农业学报》2021年第3期654-661,共8页Southwest China Journal of Agricultural Sciences

基　　金：重庆市科研院所绩效激励引导专项(cstc2018jx jl80001);重庆市技术创新与应用发展专项面上项目(cstc2019 jscx-msxmX0345)。

摘　　要：【目的】研究酉州乌羊皮肤组织ASIP基因及其编码蛋白的结构和功能。【方法】采用RACE技术对酉州乌羊皮肤组织进行扩增,获得了酉州乌羊ASIP基因的完整转录本,并结合生物信息学方法对其结构与功能进行分析。【结果】克隆后的片段通过拼接得到一个长787 bp的序列(P1),经鉴定为酉州乌羊ASIP基因的mRNA,包括209 bp的5′UTR区,402 bp的CDS区,以及176 bp的3′UTR区,预测编码133个氨基酸,为不稳定的亲水蛋白,预测到一个约22个氨基酸的信号肽。此外,酉州乌羊ASIP基因编码区发生异义替换c.496G>T,其编码的氨基酸由丙氨酸转变为缬氨酸(p.Ala96Val),该异义替换位于负责黑素皮质素受体1(MC1R)结合活性的Agouti家族保守结构域上,造成了酉州乌羊ASIP三级结构的改变。【结论】通过对酉州乌羊ASIP基因的序列特征和蛋白结构的初步分析,推测酉州乌羊皮肤组织中ASIP基因编码区发生的异义替换可能对酉州乌羊皮肤色素沉积具有影响。【Objective】The present paper aimed to study the structure and function of ASIP gene and its coding protein in Youzhoudark goat(YZD goat)skin tissues.【Method】YZD goat ASIP gene was cloned by 5′and 3′RACE from skin tissues and analyzed by the bioinformatics.【Result】A 787 bp sequence(P1)was identified as YZD goat ASIP gene including 209 bp 5′UTR,402 bp CDS,and 209 bp 3′UTR.It encoded 133 amino acids and was predicted to be an unstable hydrophilic protein,and a signal peptide of about 22 amino acids.In addition,a missense substitution(c.676 A>G)has been found in YZD goat ASIP gene,leading to the convert from the alanine to valine(p.Ala96 Val),located in the Agouti super family conserved domain.Meanwhile,the missense substitution of amino acids also caused the difference of spatial conformation of YZD goat ASIP.【Conclusion】The sequence characteristics and protein structure of YZD goat ASIP gene were analyzed and it could be concluded that the missense substitution of ASIP gene coding region in YZD that goat may have an effect on skin pigmentation.

关 键 词：酉州乌羊 皮肤 ASIP RACE克隆 

分 类 号：S826[农业科学—畜牧学]