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为研究芹菜(Apium graveolens)衔接蛋白(Adaptor protein)复合体AP-2中的μ2亚基的功能,以‘六合黄心芹’和‘美国西芹’为试验材料,采用RT-PCR方法获得AgMu2基因。序列分析表明:AP-2复合体AgMu2基因全长1 317个核苷酸,编码438个氨基酸。推测其蛋白质分子量为49.30 kD,pI为9.28。进化分析表明,来自‘美国西芹’的AgMu2亚基在植物间进化具有高度保守性,与葡萄Mu2进化关系最为接近。空间结构分析表明,从芹菜中分离的AgMu2亚基由5个螺旋和26个延伸主链构成,两个平行的β延伸主链构成的平面区域可能含有识别YXXΦ结构的位点。实时定量荧光PCR显示,芹菜中AgMu2基因主要在叶中表达,具有明显的组织特异性,同时该基因可响应低温、高温、干旱和盐胁迫等多种逆境信号,2个品种间该基因响应的时间和强度也具有显著的差异,显示了该基因功能的多样性。
In order to study the function of the μ2 subunit in AP-2, an adapter protein complex of Apium graveolens, the AgMu2 gene was obtained by RT-PCR using ’Centella roxburghii’ and ’American celery’ as experimental material. Sequence analysis showed that the AgMu2 gene of AP-2 complex was 1 317 nucleotides in length and encoded 438 amino acids. The molecular weight of the protein was estimated to be 49.30 kD and pI was 9.28. Phylogenetic analysis showed that the AgMu2 subunit from ’Celery’ was highly conserved among plants and most closely related to Mu2 evolution in grapes. Spatial structure analysis showed that the AgMu2 subunit isolated from celery consisted of 5 helices and 26 extended backbones. The planar region formed by two parallel β-extension backbones may contain sites that recognize the YXXΦ structure. Real-time quantitative PCR showed that the AgMu2 gene in celery was mainly expressed in the leaves and had obvious tissue specificity. The gene could respond to various adverse signals such as low temperature, high temperature, drought and salt stress. The response of the two genes to the gene Time and intensity also have significant differences, showing the diversity of the gene’s function.