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以华南8号木薯为材料,采用RT-PCR技术,克隆得到1个具有完整阅读框的木薯膜联蛋白基因,命名为MeAnn1(Gen Bank序列号为:KM975562),该基因CDS序列长度为951 bp,编码317个氨基酸。生物信息学分析发现,其理化性质、蛋白保守结构域及多个蛋白功能位点与已报道的植物膜联蛋白一致。进化树分析表明,木薯MeAnn1与Ptr Ann6、At Ann1、At Ann2、At Ann6和Ann Bj1具有较近的亲缘关系,具有59.9%~84%的序列相似性。细胞定位结果表明MeAnn1蛋白主要定位于细胞质和细胞核中。实时荧光定量PCR分析表明,木薯膜联蛋白基因MeAnn1的表达受多种非生物胁迫的诱导,其中低温胁迫下基因表达量上调幅度最大,为对照表达量的24倍;但对H2O2胁迫不敏感。本研究结果有助于进一步研究MeAnn1基因的功能及木薯抗逆的分子机理。
A cassava annexin gene with a complete reading frame was cloned by RT-PCR from cassava No.6 in South China and named MeAnn1 (GenBank accession number: KM975562). The length of CDS sequence was 951 bp , Encoding 317 amino acids. Bioinformatics analysis found that its physical and chemical properties, protein conserved domains and multiple protein functional sites with the reported plant annexin. Phylogenetic tree analysis showed that cassava MeAnn1 has a close genetic relationship with Ptr Ann6, At Ann1, At Ann2, At Ann6 and Ann Bj1, and has 59.9% -84% sequence similarity. The results of cell localization showed that MeAnn1 mainly localized in cytoplasm and nucleus. Real-time quantitative PCR analysis showed that the expression of MeAnn1 in Cajanus cajanon was induced by a variety of abiotic stresses. The gene expression level was up-regulated 24 times under low temperature stress, but it was not sensitive to H2O2 stress. The results of this study are helpful to further study the function of MeAnn1 gene and the molecular mechanism of cassava stress.