| 摘要: |
| 【目的】对红花油酸去饱和酶FAD2基因进行克隆和生物信息学分析，为深入研究该酶的功能提供理论依据。【方法】提取红花种子总RNA，通过RT-PCR方法进行反转录，PCR扩增得到FAD2基因全长cDNA序列。运用生物信息学方法，对红花FAD2氨基酸序列的理化性质、系统进化、蛋白定位和跨膜区域等进行预测和分析。【结果】成功克隆获得了红花FAD2基因，其开放阅读框cDNA 长1 140 bp，编码380个氨基酸，预测分子质量43.6 ku，理论等电点8.37，带有正电残基（强酸性）30个、负电残基（强碱性）34个。FAD2蛋白含有3个极度保守的His-Box，不存在明显的信号肽，存在6个跨膜结构域，与疏水区域预测结果一致。红花FAD2核苷酸的分子进化树分析显示，其与日本栽培稻的亲缘关系较近。红花FAD2氨基酸序列比对结果表明，不同植物的FAD2具有较高的同源性，同源性平均达72.94%。【结论】成功克隆了红花FAD2基因，并对其编码的蛋白进行了生物信息学分析。 |
| 关键词: 红花 油酸去饱和酶 基因克隆 同源性分析 信号肽 |
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| 基金项目:国家教育部博士点基金青年教师基金项目(20122223120002)；2014年国家大学生创新创业训练项目（201410193037）；吉林省教育厅项目（201458）；吉林省中医药管理局项目 |
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| Clone and sequence analysis of safflower oleic desaturase gene |
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WANG Wen-hui,QIANG Wei-dong,WANG Qing-man,et al
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| Abstract: |
| 【Objective】Safflower oil acid desaturase gene was cloned and bioinformatics analysis was conducted to provide a theoretical basis for further investigation of the enzyme.【Method】Total RNA of safflower seeds was extracted and cDNA was reverse transcripted by RT-PCR method.FAD2 gene was then obtained by PCR amplification.Using bioinformatics methods were then used to predict and analyze the physical and chemical properties,sequence alignment,phylogenetic,protein localization,and transmembrane region of amino acid sequence of safflower FAD2.【Result】The FAD2 gene was cloned successfully with the length of 1 140 bp,coding of 380 amino acids,the molecular mass of 43.6 ku,and the isoelectric point of 8.37.FAD2 contained 30 positive charged residues and 34 negatively charged residues.The FAD2 protein was unstable with 3 extremely conservative His Box.There existed six transmembrane domains which were consistent with the hydrophobic regions.The molecular evolution tree revealed the genetic relationship of FAD2 was closed between safflower and rice cultivars from Japan.The homology of the nucleotide sequence was 72.94%.【Conclusion】Safflower FAD2 gene was successfully cloned and bioinformatics analysis was conducted. |
| Key words: safflower oleic acid desaturase gene clone homology analysis signal peptide |
