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几丁质(N-乙酰氨基葡萄糖线性聚物)是病原真菌细胞壁的主要成分,几丁质酶通过破坏细胞新物质的沉积,激发寄主植物的抗病反应,使病原真菌细胞壁中的几丁质降解。植物几丁质酶具有广泛的生理活性。几丁质的酶基因工程成为目前抗真菌基因工程研究热点之一。本研究通过几丁质酶基因的克隆,构建原核表达载体pET-28a(+)-GchiI,利用IPTG诱导表达,经PAGE分析表明,该基因表达的蛋白分子量为34kD左右,其表达产物主要以包涵体的形式存在。抑菌圈试验发现,IPTG浓度为0.6mmol/L时抑菌效果最佳。本研究对利用基因工程技术培育抗病作物品种,提高作物抗病性具有重要参考价值。
Chitin (N-acetylglucosamine linear polymer) is the main component of the pathogenic fungal cell wall. Chitinase activates the host plant’s disease-resistance response by disrupting the deposition of new cellular material, allowing chitin in the pathogenic fungal cell wall degradation. Plant chitinase has a wide range of physiological activities. Chitinase gene engineering has become one of the hot topics in anti-fungal genetic engineering. In this study, chitinase gene cloned, the prokaryotic expression vector pET-28a (+) - GchiI was constructed and induced by IPTG. The result of PAGE showed that the molecular weight of the gene was about 34kD, The form of body exists. Inhibition zone test found that IPTG concentration of 0.6mmol / L best antibacterial effect. This study has important reference value for using genetic engineering techniques to cultivate disease-resistant crop varieties and improve crop disease resistance.