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目的:克隆耐盐碱果树滨梅的肌动蛋白基因actin,为该物种优异性状基因的功能鉴定提供内参。方法:利用一对actin简并引物克隆滨梅actin cDNA片段,对其进行序列和表达分析。结果:得到2条743 bp的cDNA片段,两条核苷酸序列相似性为82%,命名为PmAct1和PmAct2并在GenBank登录(分别为JX855160和JX855161);序列比对发现2条actin片段氨基酸序列与其他植物同源性均在96%以上;根据不同果树Actin相似性构建进化树,表明2个滨梅actin明显分为两种类型,但均与蔷薇科果树亲缘关系较密切;半定量RT-PCR表达谱发现PmAct1可能为组成型表达类型,而PmAct2特异地在根和叶组织中表达量较高。结论:首次获得了2个滨梅actin基因,为该物种其他功能基因的挖掘和表达分析奠定了基础。
OBJECTIVE: To clone the actin gene actin gene of salt-tolerant fruit tree Prunus mume, providing internal control for the functional identification of the excellent trait gene of this species. Methods: A pair of actin degenerate primers were used to clone the actin cDNA fragment and its sequence and expression analysis. RESULTS: Two 743 bp cDNA fragments were obtained. The two nucleotide sequences were 82% identical and named as PmAct1 and PmAct2, which were then registered in GenBank (JX855160 and JX855161, respectively). The amino acid sequence of the two actin fragments And more than 96% homology with other plants. Based on the Actin similarity of different fruit trees, it was found that there were two types of actin in the two species, but both were closely related to the Rosaceae fruit trees. Semi-quantitative RT- PCR showed that PmAct1 was constitutively expressed, while PmAct2 specifically expressed higher in roots and leaves. Conclusion: Two actin genes were obtained for the first time, which laid the foundation for mining and expression analysis of other functional genes in this species.