水牛ATF3基因的克隆、组织差异表达和功能初步分析  

Cloning, tissue differential expression and preliminary functional analysis of ATF3 gene in buffalo

作　　者：仝迎欣 刘婧予 张辉杨 张盈盈 周芳廷 TONG Yingxin;LIU Jingyu;ZHANG Huiyang;ZHANG Yingying;ZHOU Fangting(Colleg of Chemistry,Biology and Environment,Yuxi Normal University,Yuxi 653100,China)

机构地区：[1]玉溪师范学院化学生物与环境学院,云南玉溪653100

出　　处：《黑龙江畜牧兽医》2023年第9期60-66,132,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：云南省地方本科高校(部分)基础研究联合专项-面上项目(202101BA070001-072);云南省教育厅科学研究基金项目(2021J0675);云南省基础研究计划项目(202201AU070175);玉溪师范学院大学生创新创业训练计划项目(2021A047)。

摘　　要：为了揭示水牛激活转录因子3(ATF3)基因的结构与功能,试验克隆了水牛ATF3基因的完整编码区(CDS)序列,采用生物信息学方法初步分析其编码产物的理化特性、结构及功能,再通过实时荧光定量PCR方法检测该基因的组织差异表达情况。结果表明:水牛ATF3基因的完整CDS序列长为546 bp,编码181个氨基酸。水牛ATF3基因的氨基酸序列与普通牛、瘤牛、牦牛、山羊和绵羊的同源性在98.3%以上。在基于核苷酸和氨基酸序列构建的系统发育树中,水牛与牛科的其他物种普通牛、瘤牛、牦牛聚集在一起。水牛ATF3蛋白属于亲水性蛋白质,没有信号肽和跨膜结构,含有亮氨基拉链(bZIP)保守结构域,定位于细胞核中。ATF3蛋白的二级结构中α-螺旋、β-转角、延伸链和无规则卷曲占比分别为51.93%、8.84%、6.08%和33.15%,三级结构与人的ATF3蛋白(5vpe.2.A)有45%的相似度。ATF3蛋白具有转录调控因子活性,通过选择性和非共价结合到顺式调控区域内特定的双链基因组DNA序列中来调节基因组的转录。水牛ATF3基因在肌肉、乳腺、心脏和肾脏中的相对表达量较高,在其余组织中低表达或几乎不表达,且该基因在泌乳期水牛乳腺中的相对表达量显著高于非泌乳期水牛(P<0.05)。说明ATF3基因可能与水牛泌乳性状有关。In order to reveal the structure and function of buffalo ATF3 gene, the full sequence of the coding region in buffalo ATF3 gene was cloned, and the physicochemical properties, structure and function of the encoded product were analyzed by bioinformatics methods. In addition, the tissue differential expression of this gene was further detected by RT-qPCR. The results showed that the complete CDS length of buffalo ATF3 gene was 546 bp, encoding 181 amino acids. The amino acid sequence of buffalo ATF3 has more than 98.3% homology with that of common cattle, zebus, yak, goats and sheep. In the phylogenetic trees constructed based on both nucleotide and amino acid sequences, the buffalo clusters with the common cattle, zebus, yak, and other species of the bovine family. Buffalo ATF3 is a hydrophilic protein with no signal peptide and transmembrane region, but contains a conserved domain of bZIP, which is localized in the nucleus. The proportion of α-helix, β-turn, extended strand and random coil in the secondary structure of ATF3 protein was 51.93%, 8.84%, 6.08% and 33.15%, respectively. The tertiary structure was similar to that of human ATF3 protein(5vpe.2.A), which reached 45%. ATF3 protein has transcriptional regulator activity and regulates genome transcription by selectively and non-covalently binding to specific double-stranded genomic DNA sequences within the cis-regulatory region. The relative expression level of buffalo ATF3 gene was high in muscle, mammary gland, heart and kidney, but low or even not expressed in the remaining tissues tested. And the expression of this gene in the mammary gland of lactating buffalo was significantly higher than that in non-lactating buffalo. The results indicated that the ATF3 gene might be associated with lactation traits in buffalo.

关 键 词：水牛 激活转录因子3(ATF3) 生物信息学分析 组织差异表达 功能 

分 类 号：S823.8*3[农业科学—畜牧学] Q785[农业科学—畜牧兽医]