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根据已报道的刺甘草C4H基因序列(Gen Bank ID:D87520.1),采用RT-PCR技术克隆得到乌拉尔甘草悬浮细胞的C4H基因编码区序列,并对其进行生物信息学分析。另外,采用q-PCR方法对C4H基因受茉莉酸甲酯(Methyl jasmonate,Me JA)诱导后的表达模式进行分析。所克隆的C4H基因编码区开放阅读框(Opening reading frame,ORF)长为1 515 bp,编码504个氨基酸。q-PCR实验证明C4H基因受Me JA的诱导,在悬浮体系添加Me JA后12 h达到最高表达水平。
According to the sequence of reported C4H gene in GenBank (GenBank ID: D87520.1), the coding region of C4H gene of Ural licorice suspension cells was cloned by RT-PCR and bioinformatics analysis was performed. In addition, the expression pattern of C4H gene induced by methyl jasmonate (Me JA) was analyzed by q-PCR. The cloned C4H gene coding region open reading frame (Opening reading frame, ORF) length of 1 515 bp, encoding 504 amino acids. q-PCR experiments showed that the C4H gene was induced by Me JA and reached the highest level 12 h after the addition of Me JA to the suspension.