摘要
为探究狮头鹅的星形肌动蛋白2(ASTN2)基因单核苷酸多态性(SNPs)位点的蛋白和mRNA二级结构及生物学特性,以在同一环境下饲养的200只500日龄正处于繁殖期的健康狮头鹅为试验对象,构建DNA混合池,采用PCR扩增和直接测序技术对该基因进行单核苷酸多态性(SNPs)筛查,利用生物信息学方法从分子水平对狮头鹅ASTN2蛋白和ASTN2基因的mRNA结构进行功能预测。结果显示,在狮头鹅群体中共筛选出3个SNPs位点,分别位于外显子Exon2的71、104和146处,其中g.3682861 G>A偏离哈代-温伯格平衡(P<0.05)。狮头鹅ASTN2基因编码区序列全长3 939 bp,编码1 312个氨基酸,突变位点均未改变编码区氨基酸的编码,理论等电点为5.58,是一种酸性不稳定亲水性跨膜分泌蛋白,有信号肽存在。亚细胞定位结果显示,其分布概率为质膜(39.1%)、内质网(26.1%)、细胞核(13%)、细胞质(8.7%)、细胞壁(4.3%)和细胞架(4.3%),主要在质膜发挥生物学作用;3种特殊结构(EGF_like、MACPF、FN3),蛋白二级结构主要由α-螺旋和无规则卷曲组成。ASTN2基因突变前后mRNA二级最小自由能(MFE)、质心、含假结结构及折叠发生改变,且突变后自由能上升,结构稳定性变弱。本试验成功筛选出狮头鹅ASTN2基因的多态位点,为进一步筛选狮头鹅产蛋量候选基因的分子遗传标记奠定理论基础。
To explore the secondary structure and biological characteristics of the protein and mRNA at SNPs of the ASTN2 gene in Shitou goose, 200 healthy 500-day-old geese in the breeding period in the same feeding environment were used as the subjects. A DNA mixing pool for the animals was constructed, and the single nucleotide polymorphisms(SNPs) of the gene were screened by PCR amplification and direct sequencing, and the bioinformatics method was used to predict the protein and mRNA structure encoded by the ASTN2 gene of the geese at the molecular level. The results showed that three SNPs were screened in the Shitou goose population, which were located at 71, 104 and 146 of Exon2, respectively. Exon2-146 G>A deviated from the hardy Weinberg equilibrium(P<0.05). The total length of the coding region of the ASTN2 gene was 3939 bp, encoding 1312 amino acids, and the mutation sites did not change the coding of amino acids in the coding region. The theoretical isoelectric point was 5.58, The gene was an acidic unstable hydrophilic transmembrane secretory protein with signal peptides. The results of subcellular localization showed that the probability of its distribution was 13% in the nucleus, 26.1% in the endoplasmic reticulum, 39.1% in the plasma membrane, 4.3% in the extracellular including cell wall, 4.3% in the cell scaffold and 8.7% in the cytoplasm(8.7%). There were 3 special structures of the gene: EGF_Like, MACPF and FN3, and the secondary structure of the protein was mainly composed of α-helix and irregular curls. Before and after the mutation of the ASTN2 gene, its mRNA secondary minimum free energy( MFE),centroid,false knot,and folding were varied,and its free energy increased but the structural stability became weak. In this experiment,the polymorphic loci of the ASTN2 gene in Shitou geese were successfully screened,which laid a theoretical foundation for further screening the molecular genetic markers of candidate genes for egg production in Shitou geese.
作者
陈新企
姜平
刘瑞子
李立先
陈灿杰
贾汝敏
赵志辉
CHEN Xinqi;JIANG Ping;LIU Ruizi;LI Lixian;CHEN Canjie;JIA Rumin;ZHAO Zhihui(College of Coastal Agricultural Sciences,Guangdong Ocean University,Zhanjiang 524088,China;Guangdong Lixing Agricultural Development Co.,Ltd.,Raoping 515732,China;Chaopeng Ecological Breeding Co.,Ltd.of Chenghai District,Shantou City,Shantou 515825,China)
出处
《畜牧与兽医》
北大核心
2022年第7期7-14,共8页
Animal Husbandry & Veterinary Medicine
基金
广东省现代农业产业技术体系创新团队项目(2019KJ137,2020KJ137)
狮头鹅品种选育及健康养殖项目(B15337)
汕头市科技局现代农业重点项目(210729156901723)。
