‘红巴梨’果皮UFGT基因的克隆及表达分析

以‘巴梨’红色芽变品种‘红巴梨’果皮为材料,采用同源克隆技术和RACE结合的方法,克隆了UFGT(UDP-葡萄糖:类黄酮3-O-葡萄糖基转移酶)蛋白的部分cDNA,命名为Pc UFGT。结果表明:该cDNA片段长为1 089 bp,与苹果UFGT基因序列一致性达89%,氨基酸序列同源性为85%,含有糖基转移酶的UDPGT、COG1819和MGT等保守域。荧光实时定量PCR分析表明,该基因在‘红巴梨’幼果期表达强度约为‘巴梨’的2倍,而果实成熟期表达强度略低于‘巴梨’;该基因在‘红巴梨’果肉中不表达。

Cloning and Expression of UFGT Gene in Skin of Max Red Bartlett

By the methods of homologous clone and rapid amplification of cDNA ends (RACE),a cDNA fragment of UFGT (UDP-glucose:flavonoid 3-O-glucosyltransferase) gene was cloned from Max Red Bartlett (a red-skin sport of Bartlett),we named it as pcUFGT.The results of sequence analysis showed that this cDNA fragment includes 1 089 basepair;The similarity between this cDNA fragment and apple UFGT gene is 89%,and the glycosyltransferases superfamily conserved domain such as UDPGT,COG1819 and MGT,has been found in the amino acid sequence of the cDNA fragment cloned.By the method of real-time quantitative PCR,pcUFGT gene expressing mode in Bartlett and Max Red Bartlett was analyzed.The results showed that the express intensity in early red Bartlett was double of the express intensity in early Bartlett,but in ripe red Bartlett,the express intensity was slightly lower than that in the ripe Bartlett;Furthermore,no pcUFGT gene expression was detected in the flesh of Max Red Bartlett.