番茄APX蛋白的表达分析

Expression analysis of LeAPX

抗坏血酸过氧化物酶(APX)是植物体内重要的的抗氧化酶之一,在植物抵御外界的氧化胁迫中发挥着重要的作用。为了研究其功能特性,本研究以番茄为试材,将前期分离得到的APX基因编码区克隆至原核表达载体pET-30a中,经诱导表达纯化后,用该重组蛋白免疫小鼠制备抗体,用于Western蛋白表达分析。杂交结果表明,APX蛋白受低温、干旱的诱导,与基因在RNA水平上的表达模式一致,但APX蛋白对盐处理反应不敏感;H_2O_2处理后蛋白水平呈现出下降趋势,与基因表达变化趋势不一致,这或许是APX存在一种转录后调控的机制,有待进一步探讨。

APX is one of the important antioxidative enzymes in plant.It plays a key role in the resistance of plants to oxidative stress in the external environments.In order to study its function,we used tomato as material,APX gene which cloned before was constructed in the prokaryotic expression vector pET-30.After being induced and expressed,the fusion protein were used to prepare for antibody and Western-blot analysis.The results showed APX were induced by low temperature and PEG,in line with the patterns of gene expression,but was not induced by NaCl;After H_2O_2 spraying treatment,the level of APX appeared to decline,which display different pattern from that of the gene expression.There could be a mechanism of post-transcriptional regulation,which could be explored in future.