银杏类黄酮O–甲基转移酶基因的克隆与表达分析

 采集扬州大学银杏种质资源圃不同发育时期的银杏叶片提取RNA，经简并引物PCR和5′ RACE扩增，得到了5′端包含起始密码子在内的长为876 bp的银杏类黄酮O–甲基转移酶（flavonoid O-methyl-transferase，FOMT）基因cDNA片段，命名为GbFOMT-13，其在GenBank数据库注册号为JN711433。对所得核苷酸序列及推导的氨基酸序列进行BLAST比对分析和进化树分析，结果证实了GbFOMT-13为银杏的FOMT部分序列。半定量RT-PCR结果表明，GbFOMT-13表达水平在银杏叶片的不同发育时期差异很大，其表达模式表现为春季幼叶＞秋季老叶＞夏季成熟叶，与类黄酮总量增长速率变化趋势相似。

Cloning and Expression of Flavonoid O-methyltransferase Gene in Ginkgo biloba

Ginkgo biloba leaves were collected from Ginkgo Germplasm Resources Garden of Yangzhou University during different developmental periods,and were used to extract total RNA.After degenerate primer PCR and 5’RACE amplifying,a 876 bp cDNA sequence containing 5’end and start codon,named GbFOMT-13 was obtained,which had been embodied in GenBank database,and accession No.is JN711433.Alignment with BLAST of the nucleotide and deduced amino acid sequence and phylogenetic tree analysis showed that GbFOMT-13 was partial of FOMT gene from Ginkgo biloba.Semi-quantitative RT-PCR analysis revealed that GbFOMT-13 was developmental stage-specific and its expression level was the highest in the spring and the lowest in the summer,which is similar with the change of total flavonoid increase rate in Ginkgo biloba leaf.