中国荷斯坦奶牛铁蛋白基因3'端cDNA的克隆及序列分析
首发时间:2012-11-16
摘要:本研究旨在克隆和分析中国荷斯坦奶牛铁蛋白(FTH)基因的3'端cDNA及氨基酸序列,为该基因的功能研究和有效利用提供理论依据。运用 cDNA 末端快速扩增(RACE)技术从公犊肝脏组织中扩增出FTH基因的3'端cDNA序列,并利用生物信息学方法对该基因的氨基酸序列进行分析。结果表明,FTH基因3'端cDNA长为489 bp,编码81个氨基酸;经生物信息学分析表明,该基因编码的蛋白无N端信号肽及跨膜区,相对分子质量为9.1253 KD,理论等电点为5.19,脂肪系数为85.56,总亲水性为-0.342,其一级结构中存在4个功能结构域;二级结构元件以α-螺旋和β-旋转为主;同源序列分析表明,中国荷斯坦奶牛与原牛 FTH氨基酸的相似性最高(99%);系统进化树显示,在该基因座位上中国荷斯坦奶牛与所测物种的亲缘关系均较远。本研究有助于揭示FTH家族基因的进化历史,为对其进行深入的功能分析和利用研究提供参考。
关键词: 中国荷斯坦奶牛 铁蛋白 cDNA 末端快速扩增 克隆 序列分析
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3'end cloning and sequence analysis of a FTH gene from Chinese Holstein cattle
Abstract:This experiment was conducted to clone and analyze the 3' end cDNA sequence and amino acid sequence of FTH gene from Chinese Holstein cattle and provide the theory basis for functional studies and effective utilization. The 3' end cDNA was cloned from liver tissues by RACE, and bioinformatics method was used to analyze the amino acid sequence. The results showed the 3' end cDNA of FTH gene was 489 bp, encoded a peptide of 81 amino acid, its relative molecular weight was 9.1253 KD, isoelectric point was 5.19, fat index was 85.56, grand average of hydropathy was -0.342, and N-terminal signal peptides and transmembrane domains were not be included in this protein; Primary structure of FTH protein contain 4 functional domains, and secondary structure with α-helix and β-rotation mainly. The results obtained through homologous sequence indicated that FTH of Chinese Holstein cattle was highly similarity to Bos Taurus (99%), and the molecular evolution trees displayed that FTH of Chinese Holstein cattle had very low genetic relationship with other species determined in this research. These results provided clues for evolutionary studies of the FTH gene family, as well as served as references for further functional analysis and utilization.
Keywords: Chinese Holstein cattle FTH RACE clone sequence analysis
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