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植物中多药和毒性化合物外排(MATE)转运蛋白参与类黄酮在液泡中积累的过程。以高丛越橘品种“北卫”为试验材料,应用染色体步移和cDNA末端快速克隆技术克隆MATE转运蛋白基因的cDNA全长序列,命名为VcMATE6,序列提交到GenBank,登录号为KF875437。序列分析表明VcMATE6的cDNA全长1 557 bp,编码518个氨基酸,推测的蛋白相对分子质量为56.39 ku,理论等电点为6.12。多重序列比对显示VcMATE6与MATE型类黄酮转运蛋白有较高的同源性。聚类分析表明VcMATE6与MtMATE2、MlMTP77、AtTT12、VvAM1等一类植物典型的MATE型类黄酮转运蛋白聚为一类。VcMATE6具有2个MATE家族特有的Mat E结构域,TMHMM在线软件预测VcMATE6含有12个跨膜螺旋。实时荧光定量PCR分析表明,VcMATE6基因在越橘各器官和果实发育各阶段均有表达,但表达量存在差异,蓝果种子中表达量最高,极显著高于其他器官以及蓝果的果皮和果肉组织,其次为根,果实发育过程中VcMATE6表达量随果实成熟度表现出增加的趋势。
Multiplex and toxic compounds in plants MATE transporters are involved in the accumulation of flavonoids in vacuoles. The full-length cDNA sequence of MATE transporter gene was cloned using high-bilberry variety “Beiwei” with chromosome walking and rapid cloning of cDNA ends and named as VcMATE6. The sequence was submitted to GenBank under accession number KF875437 . Sequence analysis showed that the cDNA of VcMATE6 was 1 557 bp in length and encoded a polypeptide of 518 amino acids with a predicted relative molecular mass of 56.39 ku and a theoretical isoelectric point of 6.12. Multiple sequence alignment showed that VcMATE6 has high homology with MATE flavonoid transporter. Cluster analysis showed that VcMATE6 clustered with MATE-type flavonoid transporters, such as MtMATE2, MlMTP77, AtTT12 and VvAM1. VcMATE6 has two Mat E domains unique to the MATE family and TMHMM Online software predicts that VcMATE6 contains 12 transmembrane helices. Real-time quantitative PCR analysis showed that VcMATE6 gene was expressed in all stages of cranberry and various organs and fruit development, but the expression levels were different. The expression level of VcMATE6 was the highest in the seeds of the blueberry, which was significantly higher than that of other organs and fruits Tissue, followed by roots and fruit development VcMATE6 expression showed an increasing trend with the maturity of the fruit.