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根据已报道的植物木质素合成基因莽草酸/奎宁酸羟基肉桂酰转移酶(HCT)的序列设计简并引物,利用RACE技术,首次从柠条锦鸡儿中克隆得到HCT基因全长cDNA序列,命名为CkHCT,GenBank登录号为KC895507。该序列长度为1 750 bp,具备长度为1 305 bp的完整开放阅读框(ORF),该基因推导编码的蛋白质有434个氨基酸,预测等电点6.42,分子量48kD。序列比对后发现,柠条锦鸡儿HCT基因推导的氨基酸序列与其它植物HCT序列高度相似。系统进化分析显示,柠条锦鸡儿HCT基因与同属豆科的三叶草、苜蓿和蝶豆的HCT基因亲缘关系最近。
Degenerate primers were designed according to reported sequence of plant lignin-synthesizing gene shikimic acid / quinic acid hydroxycinnamoyltransferase (HCT). The full-length cDNA of HCT gene was cloned from Caragana korshinskii by RACE for the first time. Named CkHCT, GenBank accession number KC895507. The full-length open reading frame (ORF) was 1 750 bp in length and encoded 434 amino acids. The predicted isoelectric point was 6.42 and the molecular weight was 48 kD. Sequence alignment revealed that the amino acid sequence deduced from the HCT gene of Caragana korshinskii was highly similar to HCT sequences of other plants. Phylogenetic analysis showed that the HCT gene of Caragana korshinskii was closest to the HCT gene of the same leguminous clover, alfalfa and butterflies.