草莓FaTT10基因的克隆与表达分析

以‘红颜’草莓为研究材料,本研究采用RT-PCR技术克隆得到FaTT10基因并对其进行生物信息学分析,采用q-PCR技术分析其时空特异性表达模式。结果表明,其开放阅读框为1 704 bp,编码567个氨基酸,蛋白质分子量为62.36 kD,理论等电点为6.69。亚细胞定位预测显示,此基因定位于细胞质膜。同源性及进化树构建结果表明,草莓FaTT10与其他植物漆酶类序列同源性较高。实时荧光定量PCR分析表明,FaTT10在草莓果实不同发育时期及组织器官间均存在表达特异性:在叶、花、匍匐茎、根、茎和果实中都有表达,在小绿果实中表达量最高,在叶中表达量最低;在果实各发育阶段,FaTT10在小绿期表达量最高,随着发育时期推进,表达量逐渐降低。

Cloning and Expression Analysis of FaTT10 in Fragaria ananassa

FaTT10 gene was cloned by RT-PCR from strawberry(Fragaria ×ananassa Duch. Bebihoppe) and analyzed by bioinformatics. q RT-PCR was used to analyze the spatiotemporal specific expression pattern of FaTT10. The results showed that the ORF of FaTT10 was 1 704 bp, encoding 567 amino acids, while the molecular weight and isoelectric point of the protein were 62.36 k D and 6.69, respectively. Subcellular localization prediction showed that the gene was localized to the plasma membrane. The results of homology and phylogenetic construction showed that Fa TT10 of Fragaria ananassa had high homology with other plant laccase enzymes. The results of qRT-PCR analysis showed that the expression of FaTT10 gene was specific in Fragaria ananassa at different stages of development and in different tissues and organs. FaTT10 was expressed in leaves, flowers,stolons, roots, stems and fruits with the highest expression level in small green fruit and the lowest expression in leaves. At different stages of development, the highest expression level was during the small green fruit and the expression level decreased gradually with.the development stage.