[Objective] The clone of maize gene Ti ( Trypsin inhibitor gene Ti)and its expression after introduction was conducted, which would lay the basis of its research in future. [ Method ] The concentration and purity of total RNA, extracted from the seed of maize variety Kanghuang No. 3, were detected through the technique of electrophoresis. The maize gene Ti was amplified by means of RT-PCR system and the target genp was collected. The target gene connected with the vector PMD18-T was transferred into the E.coli competent cells, and then, (be recombined plasmid DNA was extracted and identified after its enzymatic digestion. The recover; gene Ti connected with the expression vector PCEX-4T-1 was introduced for its expression and the expression products was measured. [Results] The trypsin inhibitor appeals wilh the form of its dimer,which had also inhibition in vitro to the germination of spore- and hyphae growth of Aspergilhis flavus. [Concluuon] The important role of the trypsin inhibitor in the resistance-improving of corn to Aspergillus flavus was understood through the research on com gene Ti.%[目的]对玉米抗黄典霉基因(胰蛋白酶抑制剂基因Ti)进行克隆并诱导表达,为以后的研究打下基础.[方法]提取抗黄3号玉米种子总RNA,电泳检测其浓度及纯度,利用RT-PCR体系扩增玉米种子Ti基因,回收目的基因,将目的基因与PMD18-T载体连接并将连接产物转入大肠杆菌感受态细胞,提取重组质粒DNA进行酶切鉴定.回收Ti基因并与表达载体PGEX-4T-1连接,诱导表达并检测表达产物.[结果]胰蛋白酶抑制剂以二聚体的形式存在,体外对黄曲霉孢子的萌发和菌丝的生长同样具有抑制作用.[结论]通过对玉米Ti基因的研究,得出胰蛋白酶抑制剂在提高玉米对黄曲霉的抗性方面具有很重要的作用.
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