青海湖裸鲤foxk1基因克隆和初步功能分析

Cloning and Preliminary Function Analysis of foxk1 Gene in Gymnocypris przewalskii

为青海湖裸鲤人工扩繁和分子遗传育种研究提供理论基础,采用RACE和RT-PCR克隆青海湖裸鲤foxk1全长cDNA序列,对其进行生物信息分析和功能分析。结果表明:青海湖裸鲤foxk1全长cDNA序列为2251bp,包含1944-bp开放阅读框,编码647个氨基酸;克隆的foxk1与其它脊椎动物的氨基酸序列同源性为68.2%~94.4%,亲缘性相近;foxk1与其邻近基因gnal2、sdk1、mmd2的共线性在整个脊椎动物中高度保守;青海湖裸鲤foxk1高表达于垂体和性腺,中量表达于脑,微量表达于鳃和心脏;青海湖裸鲤foxk1在3月和6月卵巢中低表达,在9月及后期卵巢中持续高表达。

In order to provide a theoretical basis for artificial propagation and molecular genetic breeding of Gymnocypris przewalskii,the full-length cDNA of foxk1 was cloned fromG.przewalskii by using RT-PCR and RACE,and the biological information analysis and functional analysis were conducted as well.Results:The cloned foxk1 gene was 2251 bp,containing a 1944-bp open reading frame which encoded 647 amino acids.The alignment showed that cloned foxk1 was highly conserved with other vertebrates with the similarity between 68.2%-94.4%.Syntenic analysis showed that foxk1 and its adjacent gene gnal2,sdk1,mmd2 were highly conserved.RT-PCR displayed that foxk1 was expressed highly in the pituitaries and gonads,moderately in the brains,rarely in the gills and hearts.Real-time PCR analysis revealed that foxk1 was expressed lowly in March and June,highly in the ovaries in September and afterwards.