›› 2016, Vol. 43 ›› Issue (4): 928-933.doi: 10.16431/j.cnki.1671-7236.2016.04.011

• 生物技术 • 上一篇    下一篇

民猪FoxN1基因的克隆及生物信息学分析

李苓1, 张冬杰2, 汪亮2, 王鑫鑫1, 曹越1, 刘娣1,2   

  1. 1. 东北农业大学动物科技学院, 哈尔滨 150030;
    2. 黑龙江省农业科学院畜牧研究所, 哈尔滨 150086
  • 收稿日期:2015-10-19 出版日期:2016-04-20 发布日期:2016-04-27
  • 通讯作者: 刘娣 E-mail:liudi1963@163.com
  • 作者简介:李苓(1989-),女,河北沧州人,硕士,研究方向:动物遗传育种与繁殖,E-mail:zgxmsywbxh@163.com
  • 基金资助:
    国家自然科学基金项目(31301955);黑龙江省自然科学基金项目(C201312);国家生猪产业体系岗位科学家(CARS-36)

Cloning and Bioinformatics Analysis of FoxN1 Gene of Min Pig

LI Ling1, ZHANG Dong-jie2, WANG Liang2, WANG Xin-xin1, CAO Yue1, LIU Di1,2   

  1. 1. College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China;
    2. Institute of Animal Husbandry, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, China
  • Received:2015-10-19 Online:2016-04-20 Published:2016-04-27

摘要: 试验旨在对民猪FoxN1基因的结构和功能进行初步探索。利用常规PCR技术对民猪FoxN1基因进行克隆,利用生物信息学手段对所获得FoxN1基因完整编码区序列进行分析,对民猪和大白猪的FoxN1基因序列进行比对,并构建分子进化树。结果显示,民猪FoxN1基因完整编码区长1 941 bp,编码646个氨基酸,分子质量为68.64 ku,理论等电点为5.81。FoxN1蛋白位于细胞核内,不存在信号肽,不是分泌性蛋白,不存在跨膜区,在第269-347氨基酸处有1个叉头结构域,该蛋白在不同的物种间有高度的保守性。克隆发现了3种转录剪切变异体,与变异体1相比,变异体2在第124-387核苷酸位置处发生缺失,变异体3在581-582处发生GCA插入。民猪与大白猪的FoxN1基因序列相比,存在4个突变位点,其中3个是同义突变,1个是错义突变,导致第108位的氨基酸发生了改变。由分子进化树得知,民猪与偶蹄目动物亲缘关系较近。上述结果表明FoxN1基因结构复杂,对其生物信息特性的探究将为进一步揭示民猪FoxN1基因的功能提供依据。

关键词: 民猪; FoxN1基因; 基因克隆; 生物信息学分析

Abstract: This study was intended to explore the structure and function of FoxN1 genes of Min pig.Min pig FoxN1 gene was cloned by PCR, the complete coding region of FoxN1 sequence was analyzed by bioinformatics methods, the FoxN1 gene sequences of Min pig and Large White pig were compared, and phylogenetic tree was built.The results showed that Min pig FoxN1 complete gene coding region was 1 941 bp, encoding 646 amino acids, molecular weight was 68.64 ku, theoretical isoelectric point was 5.81. FoxN1 protein located within the nucleus, it was not a secreted protein, in which signal peptide was not present and there was no transmembrane area.There was a FH domain in 269 to 347 amino acid position.The protein among different species was highly conserved.Three kinds of transcription splice variants were found.Compared with variant 1, there was deletion in the nucleotide position 124 to 387 in variant 2, and GCA insertion in 581 to 582 in variant 3.Compared with Large White pig, Min pig FoxN1 gene sequences existed four mutations, of which three were synonymous mutations, one was a missense mutation, resulting in the 108th amino acid changed.The phylogenetic tree revealed a close genetic relationship between Min pig and artiodactyla animals.The results showed the genetic structure of FoxN1 gene was complex, it would provide the basis for a further reveal on Min pig FoxN1 gene's function.

Key words: Min pig; FoxN1 gene; gene cloing; bioinformatics analysis

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