苦瓜核糖体失活蛋白MAP30(Momordica charantia antiviral 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.将舍有重组表达质粒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~(-1) IPTG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 poly-clonal antibodies have been raised in rabbits immuned with the purified recombinant protein.
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