油葵HaFAD2-2基因的克隆及表达分析
发布时间：2018-12-23 09:25
【摘要】：脂肪酸脱氢酶2(fatty acid desaturase,FAD2)催化油酸生成亚油酸,是植物体内生成多不饱和脂肪酸的关键酶。根据已报道的向日葵(Helianthus annuus L.)FAD2基因序列,设计引物进行RT-PCR,克隆得到油葵FAD2-2基因全长cDNA,命名为HaFAD2-2。该基因开放阅读框为1 152bp,编码383个氨基酸,相对分子质量43.96kD,等电点为8.56。对基因组进行内含子调查发现,该基因在编码区内没有内含子。多序列比对和系统进化分析发现,FAD2-2基因编码蛋白与金盏菊(Calendula officinalis)、斑鸠菊(Vernonia galamensis)等菊科植物具有较近的亲缘关系。qRT-PCR分析表明,HaFAD2-2基因在根、茎、叶、花、子叶和未成熟种子中均有表达,且以叶中的表达量最高,未成熟种子中的表达量最低;低温(5℃、15℃)胁迫处理能显著促进该基因在根中的表达,抑制其在叶中的表达;盐胁迫(300 mmol/L NaCl)处理对其表达也具有抑制作用。该研究结果可为进一步探讨HaFAD2-2基因的功能奠定基础。
[Abstract]:Fatty acid dehydrogenase (2 (fatty acid desaturase,FAD2) catalyzes oleic acid to produce linoleic acid, which is the key enzyme to produce polyunsaturated fatty acids in plants. According to the reported sequence of sunflower (Helianthus annuus L.) FAD2 gene, a primer was designed for RT-PCR, cloning to obtain the full length cDNA, of FAD2-2 gene of oil sunflower named HaFAD2-2.. The open reading frame of the gene is 1152 BP, encoding 383 amino acids, the relative molecular weight is 43.96 kD and the isoelectric point is 8.56. An intron investigation of the genome revealed that the gene had no intron in the coding region. Multiple sequence alignment and phylogenetic analysis showed that the FAD2-2 gene encoded protein was closely related to (Calendula officinalis), (Vernonia galamensis) and other plants of Compositae. QRT-PCR analysis showed that HaFAD2-2 gene was found in roots, stems, leaves and flowers. Both cotyledons and immature seeds had the highest expression, and the lowest in immature seeds. The low temperature (5 鈩，
本文编号：2389771
[Abstract]:Fatty acid dehydrogenase (2 (fatty acid desaturase,FAD2) catalyzes oleic acid to produce linoleic acid, which is the key enzyme to produce polyunsaturated fatty acids in plants. According to the reported sequence of sunflower (Helianthus annuus L.) FAD2 gene, a primer was designed for RT-PCR, cloning to obtain the full length cDNA, of FAD2-2 gene of oil sunflower named HaFAD2-2.. The open reading frame of the gene is 1152 BP, encoding 383 amino acids, the relative molecular weight is 43.96 kD and the isoelectric point is 8.56. An intron investigation of the genome revealed that the gene had no intron in the coding region. Multiple sequence alignment and phylogenetic analysis showed that the FAD2-2 gene encoded protein was closely related to (Calendula officinalis), (Vernonia galamensis) and other plants of Compositae. QRT-PCR analysis showed that HaFAD2-2 gene was found in roots, stems, leaves and flowers. Both cotyledons and immature seeds had the highest expression, and the lowest in immature seeds. The low temperature (5 鈩，
本文编号：2389771
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