甘蓝型油菜植物防御素基因的克隆与原核表达  

Gene Cloning and Prokaryotic Expression of Brassica napus Plant Defensin

作　　者：王轶[1] 罗勤[1] 吴俊[1] 郭经宇[1] 杨毅[1] 李旭锋[1] 

机构地区：[1]四川大学生命科学学院植物遗传实验室,成都610064

出　　处：《四川大学学报（自然科学版）》2005年第4期831-834,共4页Journal of Sichuan University(Natural Science Edition)

摘　　要：根据Genbank上已知的植物防御素基因设计引物,以甘蓝型油菜中的品种”蜀杂九号”种子总RNA反转录成的cDNA为模板,进行PCR扩增,获得了243bp的片段.将该片段回收,连接到pMD18-T载体测序.将测序片段经酶切回收后克隆到GTK表达载体中,在0.1mmolIPTG诱导下表达出与理论值相符的34kD的融合蛋白条带.用GST单克隆抗体做第一抗体进行Western-blot检测,获得阳性结果.这为甘蓝型油菜植物防御素基因功能的进一步研究提供了基础.Design a couple of primers according to the known sequence of plant defensins gene from Genbank.Extract the total RNA from Shuza No. 9 of Brassica napus seeds and then revert to cDNA. The gene was amplified by PCR and cloned into pMD18-T vecter , then was digested and inserted into the prokaryotic expression GTK vector. Protein was expressed under the induction of IPTG, and a protein band with a relative molecular weight of 34 kid was obtained and was recognized by GST antibody. This should provide a powerful tool for investigating the function of Brassica napus plant defensin.

关 键 词：甘蓝型油菜 植物防御素 融合蛋白 原核表达 

分 类 号：Q78[生物学—分子生物学]