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以‘石硖’龙眼为试材,采用RT-PCR结合RACE技术成功克隆一个龙眼漆酶基因全长c DNA序列,命名为Dl Lac,NCBI登录号为KY051551。Dl Lac基因序列全长1898 bp,编码576个氨基酸,NCBI比对结果显示其与荔枝漆酶氨基酸序列同源性最高,高达94%;进化树结果显示龙眼漆酶氨基酸序列与荔枝漆酶氨基酸序列具有高度同源性。氨基酸保守序列结果表明龙眼漆酶氨基酸序列含有漆酶的3个典型保守结构域,分别为:Cu-oxidase-3、Cu-oxidase和Cu-oxidase-2。结合龙眼果实在常温、低温贮藏条件下,果皮褐变与Dl Lac的表达关系,推测Dl Lac的上调表达可能对龙眼果皮褐变起促进作用。
The full-length c DNA sequence of a longan laccase gene was cloned by RT-PCR and RACE technology, named as Dl Lac. NCBI accession number was KY051551. The Dl Lac gene was 1898 bp in length and encoded 576 amino acids. The NCBI alignment showed that the amino acid sequence of Dl Lac had the highest homology with the amino acid sequence of lychee laccase (94%). The phylogenetic tree showed that the amino acid sequence of the longan laccase and the amino acid sequence of litchi laccase Has a high degree of homology. Amino acid conserved sequence results showed that the amino acid sequence of longan laccase contains three typical conserved domains of laccase, namely Cu-oxidase-3, Cu-oxidase and Cu-oxidase-2. Based on the relationship between the browning of peel and the expression of Dl Lac at room temperature and low temperature, the up-regulation of Dl Lac may promote the browning of longan pericarp.