中国畜牧兽医

• 生物技术 •    下一篇

京海黄鸡肌细胞生成素基因的克隆和生物信息学分析

张涛1,2,张跟喜1,2,王金玉1,2,樊庆灿1,2,王文浩1,2,魏岳1,2,陈学森1,2,唐莹1,2,王永娟3   

  1. 1.扬州大学动物科学与技术学院,江苏扬州 225009;2.江苏省动物遗传繁育与分子设计重点实验室,江苏扬州 225009;3.江苏京海集团,江苏南通 226103
  • 收稿日期:2014-01-07 出版日期:2014-08-20 发布日期:2014-08-22
  • 通讯作者: 王金玉。E-mail:jywang@yzu.edu.cn
  • 作者简介:张涛(1990—),男,山东人,硕士生,研究方向:动物遗传育种与繁殖。
  • 基金资助:

    国家肉鸡产业技术体系(nycytx-42-G1-05);江苏高校优势学科建设工程;江苏省动物遗传繁育与分子设计重点实验室;江苏省高校自然科学研究(12KJB230003)。

Cloning and Bioinformatics Analysis of MyoG Gene in Jinghai Yellow Chicken

ZHANG Tao1,2, ZHANG Gen-xi1,2,WANG Jin-yu1,2, FAN Qing-can1,2, WANG Wen-hao1,2, WEI Yue1,2, CHEN Xue-sen1,2, TANG Ying1,2,WANG Yong-juan3   

  1. 1. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;2. Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design of Jiangsu Province, Yangzhou 225009, China;3. Jiangsu Jinghai Poultry Group Co., Ltd., Nantong 226103, China
  • Received:2014-01-07 Online:2014-08-20 Published:2014-08-22

摘要: 本试验根据GenBank中公布的原鸡肌细胞生成素(myogenin,MyoG)基因序列设计1对引物,采用RT-PCR方法,从京海黄鸡肌肉组织中克隆MyoG基因的编码区序列,并使用多种生物软件和在线工具对目的序列进行生物信息学分析,分析MyoG 基因与其他物种的同源性、蛋白质的理化性质、蛋白质序列的跨膜区、亚细胞定位、亲水性、潜在的磷酸化位点、保守结构域及该基因编码蛋白的二级结构和三级结构。最终获得了包括编码区在内的长754 bp的MyoG基因序列,生物信息学分析显示,京海黄鸡MyoG基因的保守性较低,与GenBank中原鸡、火鸡、鹌鹑、马、人、野猪、牛、山羊、大鼠、绵羊的同源性分别为99.7%、94.4%、92.0%、70.0%、70.0%、73.3%、69.0%、67.9%、67.8%、73.5%;氨基酸分析发现,MyoG蛋白为水溶性蛋白,分子质量为25854 u,理论等电点为5.46,不属于跨膜蛋白;亚细胞定位显示,大部分蛋白位于细胞质内,不属于分泌蛋白;预测含有6个潜在的磷酸化位点,1段碱性序列,1段HLH序列,1段低复杂度序列;二级结构以无规则卷曲为主,三级结构显示MyoG蛋白的结构域呈螺旋状。

关键词: 京海黄鸡; 肌细胞生成素; 克隆; 生物信息学分析

Abstract: Based on the published mRNA nucleotide sequence of Gallus gallus myogenin(MyoG) gene, a pair of primers were designed to clone the MyoG gene coding sequence of Jinghai Yellow chicken by RT-PCR. A variety of software and online tools were used for bioinformatic analysis of MyoG gene. Bioinformatics technology was successfully used to analyze the MyoG homology among different species, physical and chemical properties of the protein, transmembrane region of MyoG sequence, subcellular localization, hydrophilic, potential phosphorylation locus, conserved domain database, MyoG secondary structure and teriary structure. Finally, a 754 bp gene sequence was cloned, which contained CDS region, promoter region and part of the 3 ′region. Bioinformatics analysis showed that MyoG gene of Jinghai Yellow chicken was less conservative and shared 99.7%, 94.4%, 92.0%, 70.0%, 70.0%, 73.3%, 69.0%, 67.9%, 67.8% and 73.5% identity with Gallus gallus, Meleagris gallopavo, quail, horse, people, wild boar, cattle, goats, rats and sheep. The amino acid sequence analysis revealed that the MyoG gene of Jinghai Yellow chicken encoded water-soluble protein and its molecular weight was 25854 u, isoelectric point was 5.46, and did not belong to the membrane protein. Subcellular localization of MyoG was in the cytoplasm and it did not belong to the secreted protein. The MyoG protein contained six phosphorylation sites,a alkaline sequences, a HLH sequence, a low complexity sequence. The secondary structure of MyoG was mainly composed of random coil. The tertiary structure of domain area of MyoG protein showed a helix structure. The results above laid the foundation for further studies of expression regulation mechanism and function of MyoG gene in the muscle growth process.

Key words: Jinghai Yellow chicken; myogenin; cloning; bioinformatics analysis