草莓ζ-胡萝卜素脱氢酶基因ZDS的克隆及特征分析

Cloning and Characterization of ζ-carotene Desatura Gene in Fragaria ananassa Duchesne

摘要:

用RT-PCR和RACE技术从草莓(Fragaria ananassa Duchesne)果实中克隆到ζ-胡萝卜素脱氢酶基因ZDS，命名为FaZDS，其cDNA全长2148 bp，具有1个1710 bp的完整开放阅读框(ORF)，编码569个氨基酸。序列分析表明，FaZDS编码的氨基酸序列与其它植物的ZDS蛋白有很高的相似性。系统进化树分析表明，草莓与苹果的ZDS蛋白亲缘关系较近。原核表达结果表明FaZDS基因在大肠杆菌中能高效表达。用半定量RT-PCR技术进行组织表达模式分析表明，FaZDS基因在草莓的花、叶片和果实中均有表达，表达量为花＞红果＞粉红果＞白果＞青果＞叶。

Abstract:

ζ-carotene Desatura (ZDS) is one of the rate-limiting enzymes in the carotenoids biosynthetic pathway. The ZDS gene named as FaZDS was cloned from strawberry (Fragaria ananassa Duchesne) fruit using RT-PCR and RACE techniques. The results showed that the length of FaZDS cDNA was 2148 bp with an open reading frame of 1710 bp, which encoded a polypeptide of 569 amino acids. Homology analysis showed that the deduced ZDS protein was highly homologous to those from other species. Phylogenetic analysis also indicated that ZDS in strawberry was close to apple. Prokaryotic expression showed that recombinant FaZDS had highly expression in E. coli BL21. The FaZDS could express in flower, fruit and leaf by semi-quantitative RT-PCR analysis, with the order as flower > red fruit > pink fruit > white fruit > green fruit > leaf.