[1]朱 鹏,陈集成,陈 凤,等.水牛Sohlh2基因的克隆分析及其表达[J].江苏农业学报,2016,(02):399-407.[doi:10.3969/j.issn.1000-4440.2016.02.025]
　ZHU Peng,CHEN Ji-cheng,CHEN Feng,et al.Cloning and expression pattern of buffalo Sohlh2 gene[J].,2016,(02):399-407.[doi:10.3969/j.issn.1000-4440.2016.02.025]
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水牛Sohlh2基因的克隆分析及其表达()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2016年02期
页码:
399-407
栏目:
畜牧兽医·水产养殖
出版日期:
2016-03-20

文章信息/Info

Title:
Cloning and expression pattern of buffalo Sohlh2 gene
作者:
朱 鹏1; 2;  陈集成1;  陈 凤1;  苏 节1; 3;  李海洋1;  韦英明1;  杨素芳1;  石德顺1
1.广西大学亚热带生物资源保护利用国家重点实验室,广西 南宁 530004; 2.中国农业科学院广西水牛研究所,广西 南宁 530001; 3.广西医科大学医学科学实验中心,广西 南宁 530001
Author(s):
ZHU Peng1; 2;  CHEN Ji-cheng1;  CHEN Feng1;  SU Jie1; 3;  LI Hai-yang1;  WEI Ying-ming1;  YANG Su-fang1;  SHI De-shun1
1.State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530005, China; 2.Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Science, Nanning 530001, China; 3.Medical Scientific Research Centre, Guangxi Medical University, Nanning 530001, China
关键词:
水牛;  Sohlh2基因;  克隆分析;  表达
Keywords:
buffalo;  Sohlh2 gene;  cloning;  expression
分类号:
S823.8+3
DOI:
10.3969/j.issn.1000-4440.2016.02.025
文献标志码:
A
摘要:
为阐明广西沼泽型水牛Sohlh2基因在水牛不同组织中的表达规律,根据GenBank已公布的牛Sohlh2基因序列,设计其编码区(CDS)扩增引物,应用RT-PCR方法扩增克隆获得目的基因片段; 应用生物信息学方法,系统分析了水牛Sohlh2的遗传进化以及蛋白质的理化性质、二级和三级结构; 并应用RT-QPCR技术对Sohlh2基因在水牛组织(器官)的表达进行了分析。结果表明:水牛Sohlh2基因编码区全长1 281 bp,共编码426个氨基酸。多重序列比较分析结果显示水牛Sohlh2核苷酸序列与牛、绵羊、山羊、猪、马、人和小鼠相应序列的相似性分别为98%、96%、96%、85%、86%、81%和72%,系统进化树显示Sohlh2基因在不同物种以及进化的过程中具有高度保守性。Sohlh2蛋白质呈弱碱性,无信号肽,在细胞中亚定位于细胞核中,存在bHLH结构。水牛Sohlh2基因在水牛垂体、大脑、肝脏、骨骼、卵巢和睾丸等11个组织(器官)细胞中具有不同程度的表达,其中睾丸细胞中表达量最高,垂体和生殖脊细胞中表达量次之,卵丘细胞中表达量低,心脏、肺脏、肌肉细胞和颗粒细胞中不表达。
Abstract:
To identify the expression pattern of Sohlh2 gene in different tissues of Guangxi swamp buffalo, a pair of special primers for CDS of Sohlh2 gene was designed according to the released sequence of bovine Sohlh2 in GenBank. The Sohlh2 gene was amplified by RT-PCR, and its genetic evolution, protein physical and chemical properties, the secondary and tertiary structure were systemically analyzed by bio-informatics techniques. A 1 281 bp whole length coding region encoding 426 amino acids of buffalo Sohlh2 gene was cloned and sequenced. The multiple sequence alignment revealed that buffalo Sohlh2 gene shared 98%, 96%, 96%, 85%, 86%, 81% and 72% similarities in nucleotide sequence with Bos Taurus, Ovis aries, Capra hircus, Sus scrofa, Equus caballus, Homo sapiens and Mice, respectively. Phylogenetic tree analysis showed that Sohlh2 gene was highly conserved in different species during their evolution. Sohlh2 protein was weakly alkaline, signal peptide free, and was localized in nuclear. The secondary structure of protein Sohlh2 contains 20 alpha helixes, 4 beta sheets, 23 beta turns and 18 random coils. RT-qPCR results showed that the expression levels of buffalo Sohlh2 varied in 11 of 15 tested samples, with the most abundant expression in testis, followed by hypophysis and germinal ridge, and the minimal expression in cumulus cells. There was no expression in lung, heart, muscle and granular cells.

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备注/Memo

备注/Memo:
收稿日期:2015-05-04 基金项目:国家“863”计划项目(2011AA100607); 广西自然科学基金(回国重点)项目(2013GXNSFCB019001); 广西自然科学基金项目(2014GXNSFAA118114) 作者简介:朱 鹏(1985-),男,江西高安人,博士,主要从事动物遗传育种与繁殖研究。(E-mail)yijianrudi@163.com。陈集成为并列第一作者。 通讯作者:杨素芳,(E-mail)ysfang3511@163.com; 石德顺,(E-mail)ardsshi@g
更新日期/Last Update: 2016-03-20