摘要
苦瓜核糖体失活蛋白MAP30(Momordica charantiaantiviral protein 30)是苦瓜中一种具有抗病毒、抗肿瘤、抗菌等多种生物活性的蛋白质.将含有重组表达质粒pET-28(a)-MAP30的大肠杆菌BL21进行诱导表达获得了重组蛋白MAP30,对IPTG(异丙基-β-D-硫代半乳糖苷)诱导浓度、诱导时间表达条件的分析表明,工程菌经0.2mmol.L-1IPTG诱导6 h实现高效可溶性表达.利用镍离子鏊合亲和层析对重组蛋白纯化,当洗脱液中咪唑的浓度为100 mmol.L-1时,目的蛋白被洗脱的效果最好.将纯化后的重组蛋白免疫家兔,获得了特异性的多克隆抗血清.
Ribosome-inactivating proteins MAP30 in bitter melon（Momordica charantia L.） is a protein with anti-virus,anti-tumor,anti-bacterial and other biological activities.The recombinant plasmid pET-28（a）-MAP30 was transformed into E.coli BL21 and induced to express by IPTG.After induced by 0.2 mmol·L^-1 IPTG at 30 ℃ for 6 h,the fusion protein His-MAP30 was produced in high-level and soluble form.The fusion protein was purified by Ni^2＋-NTA column.The target protein was eluted efficiently when the imidazole concentration of elution buffer was 100 mmol · L^-1. The specific polyclonal antibodies have been raised in rabbits immuned with the purified recombinant protein.
出处
《河南农业大学学报》
CAS
CSCD
北大核心
2009年第5期552-556,共5页
Journal of Henan Agricultural University
基金
河南农业大学博士研究基金项目(30200274)
关键词
苦瓜
苦瓜核糖体失活蛋白
原核表达
抗体制备
bitter melon
ribosome inactivating protein MAP30
prokaryotic expression
antibody preparation
