Cloning and expression analysis of CYP83B1 from Isatis indigotica Fort
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摘要: 对菘蓝（Isatis indigotica Fort.）CYP83B1基因进行了克隆与表达模式分析。结果显示，IiCYP83B1基因全长为1652 bp，包含2个外显子和1个内含子；cDNA全长为1500 bp，编码499个氨基酸。IiCYP83B1编码的蛋白没有跨膜结构域和信号肽，主要定位于内质网膜，属于亲水性蛋白，二级结构主要由无规则卷曲螺旋和α-螺旋组成，与萝卜（Raphanus sativus Linn.）、欧洲油菜（Brassica napus L.）、甘蓝（Brassica oleracea L.）和芜菁（Brassica rapa L.）等植物的CYP83B1蛋白具有较高的同源性。qRT-PCR分析结果表明，IiCYP83B1基因在菘蓝的根、茎、叶、花和果中均有表达，且以叶中的表达量最高；在幼苗期、生长期和花期稳定表达且均显著高于萌芽期；茉莉酸甲酯（methyl jasmonate，MeJA）和葡萄糖（glucose，Glu）能够显著促进该基因的表达，而低温（4℃）和水杨酸（salicylic acid，SA）处理对其表达具有一定的抑制效应。本研究结果可为进一步探讨IiCYP83B1基因的功能提供参考。Abstract: The CYP83B1 gene of Isatis indigotica Fort. was cloned and its expression patterns were analyzed. Results showed that the length of the IiCYP83B1 gene was 1652 bp, and included two exons and one intron. The full length cDNA of IiCYP83B1 was 1500 bp, encoding a protein of 499 amino acids. IiCYP83B1 was a hydrophobic protein located in the endoplasmic reticulum, without a transmembrane domain or signal peptide. Its secondary structure mainly included alpha helixes and irregular coils. Homologous comparison illustrated that IiCYP83B1 has close relationship with Raphanus sativus Linn., Brassica napus L., Brassica oleracea L., and Brassica rapa L. qRT-PCR analysis indicated that IiCYP83B1 was expressed in root, stem, flower, and fruit, and highly expressed in leaf. It was also highly expressed in the seedling, vegetative growth, and flowering stages, compared with the germination period. Moreover, IiCYP83B1 could be induced significantly by methyl jasmonate (MeJA) and glucose (Glu), but repressed by low temperature (4℃) and salicylic acid (SA). Results in this experiment provide reference for further functional study on IiCYP83B1.
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Keywords:
- Isatis indigotica Fort. /
- CYP83B1 /
- Gene clone /
- Bioinformatics /
- Expression pattern
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