芜菁的类黄酮3'羟化酶基因克隆和UV-A诱导表达特性

Cloning and UV-A Induced Expression Trait of Flavonoid 3'-Hydroxylase Gene in Turnip(Brassica rapa L.)

用UV-A处理‘津田’芜菁和‘赤丸’芜菁块根24h后提取总RNA,以RT-PCR方法分别克隆到BrF3'H1和BrF3'H2基因。BrF3'H1和BrF3'H2的开放读码框为1536bp,均编码511个氨基酸。氨基酸序列分析显示,BrF3'H1和BrF3'H2与甘蓝型油菜F3'H的同源性达99%,在第45～476的肽段含有细胞色素P450家族基因的结构域。BrF3'H1和BrF3'H2基因有高度同源性,核苷酸序列的17个位点处有差异,推导的氨基酸序列在5个位点处有差异。Northern杂交结果显示,UV-A可以诱导BrF3'H1表达,基因的表达量与UV-A处理时间呈相关,UV-A不能诱导BrF3'H2基因表达。

The roots of ＇Tsuda＇ turnip （Brassica rapa） and ＇Yurugi Akamaru＇ turnip were irradiated with UV-A light for 24 h. Total RNAs were isolated and then BrF3＇H1 and BrF3＇H2 genes were cloned by RT-PCR method. The open reading frame （ORF） of BrF3＇H1 and BrF3＇H2 genes contained 1 536 bp encoding proteins of 511 amino acids. Amino acid sequence analysis showed that BrF3＇HI and BrF3＇H2 were 99% identitied to F3＇H of Brassica napus, and the cytochrome P450 domain was in the sequence from 45 to 476. BrF3＇H1 and BrF3＇H2 genes had high identity. The nucleotide sequence of BrF3＇H1 and BrF3＇H2 genes had 17 differences, as well as 5 differences in deduced amino acid sequence. The Northern blotting results showed that the expression of BrF3＇HI could be induced by irradiation of UV-A, and the expression of the gene was correlated with lightexposure time. The expression of BrF3＇H2 gene could not be induced by irradiation of UV-A.