重组免疫毒素hIL-2-LuffinP1的构建及表达  

Construction and expression of a recombinant immunotoxin hIL-2-Luffin P1

作　　者：王儒鹏[1] 周丽娜[1] 吴军[1] 贺伟峰[1] 易绍萱[1] 陈希伟[1] 张小容[1] 

机构地区：[1]第三军医大学西南医院全军烧伤研究所,创伤,烧伤与复合伤国家重点实验室,重庆400038

出　　处：《第三军医大学学报》2005年第9期828-830,共3页Journal of Third Military Medical University

摘　　要：目的　构建人分泌型IL 2与免疫毒素LuffinP1的重组基因原核表达载体,并对其产物进行鉴定及纯化。方法　采用基因工程原理,将IL 2 LuffinP1的重组基因片段克隆入表达载体pET2 0b(+ )中,经酶切及DNA序列测定鉴定正确,转化表达宿主菌Origami(DE3 ) pLysS中,经IPTG诱导表达含His标签的融合蛋白 重组免疫毒素hIL 2 LuffinP1。结果　成功构建了免疫毒素表达载体pET2 0b(+ ) hIL 2 LuffinP1,获得纯化的重组免疫毒素,并经Westernblot鉴定正确。结论　hIL 2 LuffinP1的成功构建,为进一步研究它在抗移植排斥中的作用奠定了基础。Objective To construct a new recombinant immunotoxin expression vector composed of human IL-2 and immunotoxin Luffin P1 gene, and to examine the inhibition effects of fusion protein hIL-2-Luffin P1 on T cells. Methods The hIL-2-Luffin P1 gene fragment was subcloned into fusion protein expression plasmid. The recombinant vector was identified by restriction endonucleases digestion and DNA sequencing. After transformed into Origami (DE3) pLysS and induced by IPTG, the protein was purified by His affinity chromatography. The effects of hIL-2-Luffin P1 on T cells were evaluated by MLR assay. Results The new recombinant immunotoxin expression vector pET20b(+)-hIL-2-Luffin P1 was constructed successfully. The fusion protein was identified by Western blot assay. Tests showed that purified IL-2-Luffin P1 had obvious inhibition effects on T cells proliferation. Conclusion hIL-2-Luffin P1 has great potentials for further research in the field of transplantation and autoimmune diseases.

关 键 词：免疫毒素 Luttin P1 WESTERN BLOT 

分 类 号：R392.11[医药卫生—免疫学] R394-33[医药卫生—基础医学]