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绿脓杆菌重组毒素分泌表达的初步研究
引用本文:彭其胜,张国利,吴广谋,朱平.绿脓杆菌重组毒素分泌表达的初步研究[J].粉末涂料与涂装,2006,19(2):155-158.
作者姓名:彭其胜  张国利  吴广谋  朱平
作者单位:吉林大学畜牧兽医学院 长春130062(彭其胜),军事医学科学院军事兽医研究所 长春130062(张国利,吴广谋,朱平)
摘    要:目的分析绿脓杆菌重组毒素的分泌表达机理。方法将所构建的6种绿脓杆菌重组毒素的分泌表达质粒,即pET-20b-IL-1023-57-PE40、pET-20b-EGF-PE40、pET-20b-LHRH-PE35、pET-20b-EGF-PE38、pET-20b-MSH-PE40和pET-20b-LHRH-PE40分别转化至Rosetta(DE3)中,进行IPTG诱导表达。结果只有LHRH-PE40和LHRH-PE35能分泌表达。LHRH-PE35表达产物经Westernblot检测,可被特异抗体识别。经DNAStar和ANThewin蛋白分析软件对6种PE重组毒素性质进行比较分析发现,并不是所有PE重组毒素融合信号肽序列后,就能分泌表达,其中导向部分的性质决定了PE重组毒素是否分泌表达。结论对绿脓杆菌重组毒素的分泌机理已有初步了解。

关 键 词:绿脓杆菌  毒素  分泌表达
收稿时间:2005-06-30
修稿时间:2005年6月30日

Secretory Expression of Recombinant Pseudomonas Extoxin
PENG Qi-sheng, ZHANG Guo-li, WU Guang-mou, et al.Secretory Expression of Recombinant Pseudomonas Extoxin[J].Chinese Journal of Biologicals,2006,19(2):155-158.
Authors:PENG Qi-sheng  ZHANG Guo-li  WU Guang-mou  
Affiliation:College of Veterinary, Jilin University, Changchun 130062, China
Abstract:Objective To analyze the mechanism of secretory expression of recombinant pseudomonas extoxin.Methods Transform the six constructed recombinant plasmids expressing pseukomonas extoxin,i.e.pET-20b-IL-10_ 23-57 -PE40,pET-20b-EGF-PE40,pET-20b-LHRH-PE35,pET-20b-EGF-PE38,pET-20b-MSH-PE40 and pET-20b-LHRH-PE40,into E.coli Rosetta(DE3) for expression under induction of IPTG.Identify the expressed product by SDS-PAGE and Western blot.Results Only LHRH-PE40 and LHRH-PE35 were expressed.Western blot showed that the expressed product of pET-20b-LHRH-PE35 was recognized by the specific antibody against LHRH-PE40.The analysis of the six kinds of recombinant pseudomonas extoxin by DNAStar and ANThewin software showed that not all the recombinant pseudomonas extoxin fused with signal peptide sequence could expressed in a form of seretory protein.The secretory expression of recombinant pseudomonas extoxin was determined by the property of leader signal.Conclusion The mechanism of secretory expression of recombinant pseudomonas extoxin was preliminarily studied.
Keywords:Pseudomonas  Extoxin  Secretory expression
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