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首页> 外文期刊>Chinese science bulletin >Inducible expression pattern of rice Bowman-Birk inhibitor gene Os WIP1-2 and its protease inhibitory activity
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Inducible expression pattern of rice Bowman-Birk inhibitor gene Os WIP1-2 and its protease inhibitory activity

机译:水稻Bowman-Birk抑制剂基因Os WIP1-2的诱导表达模式及其蛋白酶抑制活性。

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摘要

The WIP1-2 gene was cloned from rice. It belongs to the Bowman-Birk inhibitor gene family. Northern blot showed that expression of this gene was induced by wounding and jasmonic acid (JA). It indicates that the Os WIP1 gene plays an important role in the rice defense system. The Os WIP1-2 was cloned into pET28a and expressed in E. coli. Its expressed product was purified in the form of fusion protein and tested for the inhibitory activities against trypsin and chymotrypsin. It was found that the fusion protein could inhibit chymotrypsin, but not trypsin. It was also found that the His tag at its C-terminal affected its inhibitory activity significantly. The fusion protein with a natural C-terminal had the inhibitory activity, while no inhibitory activity was detected in the fusion protein with a (His)_6-tag at its C-terminal. This implies that extra amino acid residues at the C-terminal of Os WIP1-2 may interfere with its correct folding. The inhkjibitory assay indicated that the members of rice Bowman-Birk inhibitor gene family probably differentiated both in their structure and function.
机译:从水稻克隆了WIP1-2基因。它属于Bowman-Birk抑制剂基因家族。 Northern印迹显示该基因的表达是由创伤和茉莉酸(JA)诱导的。这表明Os WIP1基因在水稻防御系统中起重要作用。 Os WIP1-2被克隆到pET28a中并在大肠杆菌中表达。其表达的产物以融合蛋白的形式纯化,并测试其对胰蛋白酶和胰凝乳蛋白酶的抑制活性。发现融合蛋白可以抑制胰凝乳蛋白酶,但不能抑制胰蛋白酶。还发现在其C末端的His标签显着影响了其抑制活性。具有天然C末端的融合蛋白具有抑制活性,而在其C末端具有(His)-6标签的融合蛋白中未检测到抑制活性。这意味着Os WIP1-2的C端多余的氨基酸残基可能会干扰其正确折叠。 inhkjibitory分析表明,水稻Bowman-Birk抑制剂基因家族的成员可能在结构和功能上都有差异。

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