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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 sequence the chalcone synthase (CHS) gene of Ampelopsis grossedentata. Methods A pair of primers was designed based on the conserved sequences of cloned plant genes. The total RNA of Gramineae was used as template. The sequence of CHS gene was amplified by RT-PCR and ligated into pMD18-T Simple vector. The positive clones were analyzed by PCR After the test sequencing. A 1 173 bp sequence was obtained. Sequence analysis showed that the fragment encoded 390 amino acids, with a homology of more than 67.9% with the amino acid sequences of other higher plants. Conclusion The CHS gene was cloned for the first time from Grape Snakehead, which laid the foundation for efficient use of this gene.