亚麻纤维素合酶LuCesA8基因克隆与表达分析

Cloning and expression analysis of the cellulose synthase gene LuCesA8 in flax

以亚麻快速生长期茎部组织总RNA为模板,通过反转录PCR克隆纤维素合酶基因Lu Ce s A8(基因ID:Lus10029245)全长开放读码框序列。序列分析结果表明,开放读码框全长2 967 bp,共编码988个氨基酸。通过多氨基酸序列比对发现,该蛋白与麻风树Ces A8蛋白亲缘关系最近,相似度达87%,与胡杨、可可树、雷蒙德氏棉及芝麻Ces A8蛋白氨基酸序列相似性分别为86%、85%、85%、81%。荧光定量PCR结果表明,该基因在亚麻不同发育阶段表达水平存在显著差异,快速生长期表达量最高,花期和绿熟期次之,苗期最低。研究为揭示Lu Ce s A8基因在次生细胞壁纤维素合成中的作用奠定基础。

The total RNA extracted from flax stems at the stage of rapid growth was used for RT-PCR,then the full- length Open Reading Frame（ORF） sequence of Lu Ces A8（Gene ID： Lus10029245） was amplified. The sequence analysis showed that the full- length ORF of Lu Ces A8 consisted of 2 967 bp,encoding 988 amino acids. Homology analysis of the deduced amino acid showed 86%, 85%, 85%, 81%identity to Ces A8 from Populus trichocarpa, Theobroma cacao, Gossypium raimondii and Sesamum indicum, respectively. The q RT- PCR result revealed that the expression level of the gene was low at seedling stage, and relatively higher at flowering and maturity stage, this gene expressed the highest level at the stage of rapid growth. This study provided a foundation for further research of LuC esA 8 biological function in the process of cellulose synthesis of the secondary cell wall.