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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 was extracted from the stem tissue of flax in the rapid growth stage. The full-length open reading frame (ORF) of the cellulase gene Lu Ce s A8 (Gene ID: Lus10029245) was cloned by reverse transcription PCR. Sequence analysis showed that the open reading frame was 2 967 bp in length, encoding a total of 988 amino acids. The amino acid sequence similarity of Ces A8 protein from Jatropha curcas was 87%, and the amino acid sequence similarity with Ces A8 protein of Jatropha curcas, Cacao tree, Raymond cotton and Sesame was 86% , 85%, 85%, 81%. Fluorescent quantitative PCR results showed that there was a significant difference in the expression level of this gene at different developmental stages of flax, with the highest expression in rapid growth phase, followed by the flowering and green ripening stages, and the lowest in seedling stage. The research laid the foundation for revealing the role of Lu Ce s A8 gene in secondary cell wall cellulose synthesis.