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目的对阳春砂姜黄素生物合成途径的关键酶二酮辅酶A合成酶(diketide CoAsynthase,DCS)基因的编码cDNA序列进行克隆,为研究阳春砂姜黄素生物合成与基因调控奠定基础。方法结合阳春砂的转录组注释,根据编码区序列设计引物,通过PCR方法克隆阳春砂DCS基因的编码区cDNA。结果 PCR扩增了一个长1 170 bp的基因片段,该片段编码由389个氨基酸组成的DCS。同源性比对结果显示基因编码蛋白与姜黄的DCS具有氨基酸一致性达96%,与水仙等植物的查耳酮合酶(chalcone synthase,CHS)氨基酸一致性达66%~69%。进化树分析结果显示,与姜黄Curcumae Longae具有较近的亲缘关系。结论首次从阳春砂中克隆DCS基因获得其编码区序列,为分析基因表达特性及其在姜黄素生物合成中的功能奠定基础。
Objective To clone cDNA encoding the key enzyme of diketide CoAsynthase (DCS) gene in curcumin biosynthesis pathway, and lay the foundation for the study on curcumin biosynthesis and gene regulation. Methods According to the transcriptome annotation of YE Chun-sa, primers were designed according to the sequence of coding region, and the coding region cDNA of YE-CH4 DCS gene was cloned by PCR. Results A gene fragment of 1 170 bp was amplified by PCR. The fragment encoded 389 amino acids of DCS. Homology comparison showed that the amino acid identity of gene encoding protein with turmeric DCS was 96%, and the amino acid identity with chalcone synthase (CHS) of plants such as narcissus was 66% -69%. Phylogenetic tree analysis showed that Curcumae Longae had a close genetic relationship. Conclusion For the first time, we cloned the DCS gene from P. euphratica and obtained its coding region sequence, which laid the foundation for the analysis of gene expression characteristics and its function in curcumin biosynthesis.