王儒鹏,吴军,袁军,解志杰,贺伟峰,易绍萱,张小容
WANG Ru-peng; WU Jun; YUAN Jun; HE Wei-feng; YI Shao-xuan; ZHANG Xiao-rong(Burn research insititute of Southwest Hospital;The Third Military Medical University;

• 1:中国人民解放军第三军医大学西南医院全军烧伤研究所
• 2:创伤烧伤复合伤国家重点实验室
• 3:中国人民解放军第三军医大学西南医院全军烧伤研究所
• 4:创伤烧伤复合伤国家重点实验室

摘要(Abstract)：

目的: 构建免疫毒素LuffinP1的基因原核表达载体,获得免疫毒素LuffinP1,并对其进行鉴定及纯化。方法:设计寡聚核苷酸片段连接延长合成全长LuffinP1, PCR扩增后将其克隆入表达载体pET20b( +)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诱导表达蛋白LuffinP1,Westernblot鉴定。结果:成功构建了免疫毒素表达载体pET20b( +) -LuffinP1,获得纯化的免疫毒素,并经Westernblot鉴定正确。结论:通过基因工程成功表达免疫毒素LuffinP1,简化了其制备过程,有利于对其进一步研究应用。
Objective: To construct an immunotoxin Luffin P1 expression vector, then to obtain the immunotoxin Luffin P1 after its expression and purification. Methods: To synthesize the full Luffin P1 gene by ligating and extending of the designed oligo nucleotides. The Luffin P1 gene was subcloned into expression plasmid. The recombinant vector was identified by digestion with restriction endonucleases and DNA sequencing. After the gene was transferred into Origami (DE3) pLysS and induced by IPTG, the protein was expressed and purified by His affinity chromatography, and then was identified with western blot assay. Results: The immunotoxin expression vector pET20b(+)-Luffin P1 was constructed successfully. The purified protein that obtained was confirmed correct. Conclusion: The results show it is a convenient way to obtain Luffin P1 through gene engineering. It can benefit further research about the function of Luffin P1.

关键词(KeyWords)： 免疫毒素类;Luffin P1;Westernblot;核糖体蛋白质类
immunotoxins; Luffin P1; Western blot; ribosomal proteins

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金面上项目(30200261)

作者(Authors): 王儒鹏,吴军,袁军,解志杰,贺伟峰,易绍萱,张小容
WANG Ru-peng; WU Jun; YUAN Jun; HE Wei-feng; YI Shao-xuan; ZHANG Xiao-rong(Burn research insititute of Southwest Hospital;The Third Military Medical University;

DOI: 10.19367/j.cnki.1000-2707.2005.02.004

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