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目的对党参肌动蛋白(actin)基因进行克隆及序列分析。方法根据已经克隆的植物actin基因的保守序列设计一对简并性引物,以党参根部总RNA为模板,采用RT-PCR的方法扩增actin基因片段并连接到pMD19-T Simple载体上,阳性克隆经PCR检测后进行测序。结果得到一段603 bp的序列,序列分析表明,该片段编码200个氨基酸,与高等植物actin基因核苷酸序列同源性在78%以上,与其他肌动蛋白氨基酸序列同源性达90%以上。结论首次从党参中克隆出了actin基因,为有效利用该基因奠定了基础。
Objective To clone and sequence the actin gene of Codonopsis pilosula. Methods A pair of degenerate primers was designed according to the conserved sequence of the actin gene of cloned plant. The cDNA of actin gene was amplified by RT-PCR using the total RNA of Corydalis as a template. The positive clones were cloned into pMD19-T Simple vector After PCR detection sequencing. A 603 bp sequence was obtained. Sequence analysis showed that the fragment encoded 200 amino acids. The nucleotide sequence of the actin gene was higher than 78%. The homology of the deduced amino acid sequence with other actin amino acids was over 90% . Conclusion The actin gene was cloned from Codonopsis for the first time, which laid the foundation for efficient use of this gene.