龙眼胚胎F3H基因的cDNA克隆及序列分析

 采用IEF-SDS-PAGE双向电泳技术，对‘立冬本'龙眼合子胚发育不同时期的蛋白质进行分离，通过MALDI-TOF-TOF鉴定到其中一个上调差异表达的蛋白为黄烷酮-3-羟化酶 (flavanone 3-hydroxylase，F3H)。以‘立冬本’龙眼胚胎cDNA为模板，采用RT-PCR和RACE技术，获得F3H基因cDNA 1 404 bp全长序列。序列分析表明，F3H基因共编码365个氨基酸，其cDNA序列与柑橘、苹果、棉花等F3H基因的同源性均高达80%以上，该基因在GenBank中登录号为EF468104。

Cloning and Sequence Analysis of F3H Gene cDNA from Longan Embryo

The protein of 'Lidongben' longan zygotic embryos during different development stages were separated and compared by IEF-SDS-PAGE technique. Flavanone 3-hydroxylase was one of differential proteins identified by MALDI-TOF-TOF/MS. Then F3H gene cDNA sequence was cloned from longan zygotic embryos using RT-PCR and RACE technique. A 1 404 bp full-length cDNA sequence was obtained. Analysis of F3H gene cDNA indicated that it encoded a peptide containing 365 amino acids. The nucleotide sequence comparison with the F3H gene of Citrus sinensis, Gossypium hirsutum and Pyrus communi showed that identity was all higher than 80%. The sequence was accepted and released by GenBank (Accession number:EF468104)。