显齿蛇葡萄查耳酮合成酶基因cDNA克隆及蛋白质序列分析

cDNA cloning and protein sequence analysis of chalcone synthase gene in leaves of Ampelopsis grossedentata

目的对显齿蛇葡萄Ampelopsis grossedentata查耳酮合成酶(CHS)基因进行克隆及序列分析。方法根据已经克隆的植物基因的保守序列设计一对引物,以显齿蛇葡萄总RNA为模板,采用RT-PCR的方法扩增CHS基因序列并连接到pMD18-T Simple载体上,阳性克隆经PCR检测后进行测序。结果得到一段1 173 bp的序列,序列分析表明,该片段编码390个氨基酸,与其他高等植物CHS基因氨基酸序列同源性在67.9%以上。结论首次从显齿蛇葡萄中克隆了CHS基因,为有效利用该基因奠定了基础。

Objective To clone and analyze the chalcone synthase （CHS） gene sequence from the leaves of Ampelopsis grossedentata. Methods A pair of primers were designed on the basis of the conserved sequences of the cloned plant CHS gene. Using total RNA in the leaves of A. grossedentata as template, the sequence of CHS gene was cloned by reverse transcription polymerase chain reaction （RT-PCR） and then the gene was ligated with pMD18-T Simple vector. The positive clone was sequenced after the identification of clone by PCR. Results A fragment of 1 173 bp was cloned. The sequence analysis indicated that the fragment encoded 390 amino acids and shared sequence homology of more than 67.9% with CHS gene sequences from other higher plants. Conclusion It is the first report that a novel CHS gene is cloned from the leaves ofA. grossedentata. This work lays a foundation for the effective application ofCHS gene.