Molecular cloning and expression analysis of four actin genes (MiACT) from mango
Molecular cloning and expression analysis of four actin genes (MiACT) from mango
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DOI：
10.1007/s10535-012-0278-9
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发表时间：
2013-06-01
影响因子：
1.5
通讯作者：
Ou, S. -J.
中科院分区：
文献类型：
--
作者：
Luo, C.;He, X. -H.;Ou, S. -J.
肌动蛋白是真核细胞中最丰富的蛋白质，是控制细胞形态和运动的关键细胞骨架成分。本研究通过同源克隆的方法从芒果中分离到4个MiACT基因，分别命名为MiACT 1、MiACT 4、MiACT 7和MiACT 9。序列比对和系统发育分析表明，芒果的4个MiACT基因在核苷酸和氨基酸水平上高度相似。所有四种MiACT蛋白与其他物种的已知肌动蛋白具有高度的相似性。逆转录聚合酶链反应显示，四个MiACT基因组成型和稳定表达的所有器官测试。植物生长调节剂的施用和4种胁迫处理对MiACT 4、MiACT 7和MiACT 9的表达有显著影响，而MiACT 1的表达无响应。相反，四个MiACT基因的表达谱不受昼夜节律的调节。此外，MiACT 1的表达不受重金属处理的影响，且在采后不同时期，无论处理与否，MiACT 1的转录水平都相当稳定。我们的研究结果表明，这四个肌动蛋白基因在芒果的整个生命周期中发挥着重要作用，组成型和稳定表达的MiACT 1是最好的候选人作为芒果差异基因表达分析的内标。
Actin is the most abundant protein in eukaryotic cells and is a key cytoskeletal component controlling cell morphology and motility. In this study, four MiACT genes were isolated from mango by homological cloning and designated as MiACT1, MiACT4, MiACT7, and MiACT9. Sequence alignments and phylogenetic analysis demonstrated that the four MiACT genes of mango were highly similar to each other at the nucleotide and amino acid levels. All of four MiACT proteins showed high similarity to the known actin proteins from other species. Reverse transcription polymerase chain reaction revealed that the four MiACT genes were constitutively and stably expressed in all organs tested. Application of plant growth regulators and four stress treatments had a remarkable effect on the expression of MiACT4, MiACT7 and MiACT9, whereas expression of MiACT1 was unresponsive. In contrast, the expression profiles of the four MiACT genes were not regulated by diurnal rhythms. Moreover, the expression of MiACT1 was not affected by heavy metal treatments and the transcript level of MiACT1 was rather stable in different days during the post-harvest period either under treatment or not. Our results suggest that the four actin genes play important roles throughout the entire life cycle of mango; the constitutively and stably expressed MiACT1 is the best candidate as an internal standard for differential gene expression analysis in mango.