诸葛菜植物防御素基因克隆与原核表达研究  

Preliminary study of gene cloning and prokaryotic expression of plant defensin in Orychophragmus violaceus

作　　者：钱磊[1] 卢永波[1] 王洪斌[1] 刘施佳[1] 严碧云[1] 张亮[1] 李旭锋[1] 杨毅[1] 吴俊[1] 

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都610064

出　　处：《四川大学学报（自然科学版）》2009年第3期791-794,共4页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然科学基金(30671165)

摘　　要：以从正在萌发的诸葛菜种子中提取总RNA反转录成的cDNA作为模板,进行PCR扩增,获得了160 bp的植物防御素cDNA片段。然后将该片段克隆到pMD18-T栽体。经测序后将片段酶切回收,再克隆到GTK原核表达载体中。经0.1 mmol/L IPTG诱导表达,获得了与理论值相符的32 kD的融合蛋白质条带。用GST单克隆抗体做第一抗体进行Western-Blot检测,获得阳性结果。这为诸葛菜植物防御素基因功能的研究提供了基础。The total RNA was extracted from the seeds of Orychophragmus violaceus. The cDNA （160 bp） of plant defensin was amplified by PCR with the reverted cDNA as template and cloned into pMD18- T vector. After confirmed by sequencing, the cDNA of plant defensin was inserted into GTK vector for prokaryotic expression. Under the induction of ITPG, a protein band with a relative molecular weight of 32 kD was observed and detected by GST antibody. The results of the study provide valuable data for the reseach of the plant defensin gene from O.violaceus.

关 键 词：诸葛菜 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]