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目的对当归肌动蛋白(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,以当归根部总RNA为模板,采用RT-PCR方法扩增Actin基因片段并连接到pMD19-T载体上,阳性克隆经PCR检测后进行测序。结果得到一段598 bp的序列,序列分析表明,该片段编码198个氨基酸,与高等植物Actin基因核苷酸序列同源性在83%以上,与其他肌动蛋白氨基酸序列同源性达94%以上。结论首次从当归中克隆出了Actin基因,为有效利用该基因奠定了基础。
Objective To clone and sequence the Actin gene from Angelica sinensis. Methods A pair of degenerate primers was designed according to the conserved sequences of the cloned Actin gene. The total RNA was used as a template to amplify the Actin gene fragment by RT-PCR and ligated into pMD19-T vector. The positive clones were analyzed by PCR After the test sequencing. A 598 bp sequence was obtained. Sequence analysis showed that the fragment encoded 198 amino acids. The nucleotide sequence of Actin gene was higher than 83%. The sequence identity with other actin amino acids was over 94% . Conclusion The Actin gene was cloned from Angelica sinensis for the first time, which laid the foundation for the effective use of this gene.