摘要
目的:探讨分离纯化苦瓜种子核糖体失活蛋白(RIP)的方法,并研究利用纯化后的RIP抑制烟草花叶病毒(TMV)的活性。方法:用盐溶液抽提总蛋白后,经30%～90%的(NH4)2SO4分级沉淀,制成粗提液。用CMSepharoseFastFlow离子交换层析结合SephadexG-75凝胶柱分离纯化RIP。结果与结论:经SDS-PAGE检测及RNAN-糖苷酶活性鉴定,确定最终得到的蛋白为苦瓜RIP;所纯化的RIP对感染黄瓜和番茄的TMV有明显的抑制作用,但RIP的浓度并不与其抑制TMV的活性呈正相关。
Objective： To find out the purification method of the balsam pear（Momordica charantia） ribosome inactivating proteins（RIP） and to study the activity of RIP in restraining TMV. Methods： The total protein isolated from the seeds by extraction with NaC1 solution were made into crude extracts through 30%-90% ammonium sulfate fractional precipitation. Through ion exchange chromatography of CM Sepharose Fast Flow and the gel chromatography of Sephadex G-75, the RIP of balsam pear was obtained. Results ＆ Conclusion： By the SDS-PAGE test and the RNA N-glycosidase experiment the protein about 30 kD after purification was proved of RIP. The study showed that the purified RIP could clearly restrain the activity of the TMV which had infected the host of cucumber and tomato, but the concentration of the RIP was not positively correlative with its activity of restraining TMV.
出处
《生物技术通讯》
CAS
2007年第2期230-232,共3页
Letters in Biotechnology
基金
曲阜师范大学科研启动基金
关键词
苦瓜
核糖体失活蛋白
纯化
烟草花叶病毒
抑制活性
Momordica charantia
ribosome inactivating protein
purification
activity of restraining
TMV
