›› 2016, Vol. 43 ›› Issue (2): 311-318.doi: 10.16431/j.cnki.1671-7236.2016.02.004

• 生物技术 • 上一篇    下一篇

MARK2、CREB5基因的克隆和生物信息学分析

岳英伟, 王鑫, 杜淼, 马文芝, 郭宏   

  1. 天津农学院动物科学与动物医学学院, 天津 300384
  • 收稿日期:2015-07-29 出版日期:2016-02-20 发布日期:2016-03-02
  • 通讯作者: 马文芝, 郭宏 E-mail:mawenzhi1126@163.com;guohong64@163.com
  • 作者简介:岳英伟(1992-),天津人,女,硕士生,研究方向:牛分子育种肌肉发生机理,E-mail:yueyingwei2015@163.com
  • 基金资助:
    国家转基因重大专项:高产转基因肉牛新品种培育(2014ZX08007—002)

Cloning and Bioinformatics Analysis of MARK2 and CREB5 Genes in Cattle

YUE Ying-wei, WANG Xin, DU Miao, MA Wen-zhi, GUO Hong   

  1. College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300384, China
  • Received:2015-07-29 Online:2016-02-20 Published:2016-03-02

摘要: 试验旨在对牛MARK2、CREB5基因进行克隆和生物信息学分析.首先在GenBank数据库中利用人MARK2和CREB5基因序列比对获得牛的表达序列标签(ESTs)的序列,进行引物设计并利用RT-PCR方法从牛肩胛骨肌肉中克隆出MARK2和CREB5基因的cDNA序列,补足空缺部分,最终获得牛MARK2和CREB5基因.通过对牛MARK2和CREB5基因的生物学特性进行分析可知:牛MARK2基因的编码区长为2 076 bp,编码691个氨基酸,CREB5基因的编码区长1 527 bp,编码508个氨基酸;经蛋白质理化性质预测MARK2和CREB5蛋白均为亲水性蛋白;MARK2和CREB5蛋白的磷酸化位点均分布于苏氨酸(Thr)、丝氨酸(Ser)和酪氨酸(Tyr)残基上;MARK2蛋白与CREB5蛋白的二级结构预测以无规则卷曲为主;亚细胞定位预测MARK2蛋白主要分布在细胞质中,CREB5蛋白主要分布在细胞核中,两者均不属于分泌型蛋白;MARK2蛋白保守结构域预测含1个S_TKc和1个UBA功能结构域,CREB5蛋白保守结构域预测含有1个ZnF_C2H2、1个BRLZ功能结构域和5个低复杂度区域且二者均不具有跨膜结构.

关键词: 克隆; MARK2基因; CREB5基因; 生物信息学

Abstract: This research aimed to clone and analyze the biological characteristics of MARK2 and CREB5 genes in cattle.In the GenBank database,MARK2 and CREB5 genes of human were chosen to blast and expressed sequence tags (ESTs) of cattle were got.Specific primers were designed and RT-PCR was used to clone the cDNA sequence from the scapula muscle of cattle with vacancy parts making up,and then MARK2 and CREB5 genes were successfully cloned.The results showed that MARK2 gene coding region was 2 076 bp,which encoded 691 amino acids.CREB5 gene coding region was 1 527 bp,which encoded 508 amino acids.The physical and chemical properties of protein prediction showed that MARK2 and CREB5 proteins were hydrophilic protein;The phosphorylation sites in threonine (Thr),serine (Ser) and tyrosine (Tyr) residues;The secondary structure of MARK2 and CREB5 proteins were maily composed of random coil and layed a foundation for the tertiary structure;Protein subcellular localization prediction showed that MARK2 and CREB5 proteins did not belong to the secretory protein;MARK2 protein contained a S_TKc domain and an UBA function structure,CREB5 protein contained a ZnF_C2H2 function structure,a BRLZ function structure and five low complexity regions and MARK2 and CREB5 proteins had no transmembrane structure.

Key words: clone; MARK2 gene; CREB5 gene; bioinformatics

中图分类号: