抗肿瘤蛋白Onconase的原核细胞表达及细胞毒性检测  

EFFECTIVE EXPRESSION OF RIBONUCLEASE-ONCONASE IN ESCHERICHIA COLI AND ASSAYING ITS CYTOTOXIC POTENTIAL

作　　者：许国峰[1] 王庆诚 龚兴国[1] 费俭 

机构地区：[1]浙江大学生命科学院,杭州310027 [2]南方模式动物中心,上海201203

出　　处：《实验生物学报》2004年第3期227-231,共5页Acta Biologiae Experimentalis Sinica

摘　　要：本文主要利用pET22b(+)表达载体在大肠杆菌中表达一种在两栖类蛙(Rana pipiens)卵细胞内存在的核糖核酸酶—Onconase。通过包涵体复性、蛋白纯化等程序最终获得与天然蛋白活性相似的重组蛋白,并检测Onconase 对源于皮肤T 细胞淋巴瘤的Hut-78肿瘤细胞的毒性(IC_(50)=0.5μmol/L),证明Onconase 用于抗淋巴瘤的可行性。Onconase,a ribonuclease purified from Rana pipiens oocytes,has cytotoxie activityagainst several tumor cell lines in vitro.With the characteristic of containing four pairs of disul-fide bonds internal and N terminal sequence attributing mostly to its biological functions,it isdifficult to obtain the active Onconase from Escherichia coli expression system with normal strat-egy.Here,we fused the cDNA coding for Onconase in frame with the pelB signal sequence inpET22b(+) expression vector.Onconase can be effectively expressed in inclusion body without ad-ditional residues at N terminal under the proper inducing condition.After refolding and purifyingprocess,we can get the recombined Onconase which has a similar rihonuclease activity as theone isolated from oocytes.The recombined Onconase has a pronounced effect on proliferation ofHut-78 cells (IC_(50)=0.5μmol/L).

关 键 词：抗肿瘤蛋白 ONCONASE 表达 细胞毒性 核酸酶 淋巴瘤 

分 类 号：R73-36[医药卫生—肿瘤]