%0 Journal Article
%T Improved expression of immunotoxin DT386-GMCSF in E.coli<br>提高免疫毒素DT386-GMCSF在E.coli中表达量的研究
%A WANG Jian-wei
%A OUYANG-Jing
%A CUI Ting
%A QU Jian-guo
%A HUNG Tao
%A <br>王健伟
%A 欧阳晶
%A 崔婷
%A 屈建国
%A 洪涛
%J 中国生物工程杂志
%D 2007
%I 
%X The development of immunotoxin DT386-GMCSF, a fusion protein which bears the N-terminal 386 amino acids of diphtheria toxin and human granulocyte-macrophage colony-stimulating factor (GM-CSF) and targets the GM-CSF receptor (GM-CSFR), has provided a promising alternative therapy to the acute myeloid leukemia (AML). However, the poor expression of the protein in E.coli is still a bottleneck which limits the industrial production. To identify the critical down-regulating factors on the expression of DT386-GMCSF, a series of truncated mutants of DT386-GMCSF at the C-terminal of GM-CSF were generated and expressed in E.coli. The results showed that the encoding sequences for the L114 of the GM-CSF dramatically impact the expression of DT386-GMCSF. On this basis, a serial of mutants integrating amino acid substitutes were generated. The results revealed that the expression level of the mutant DF123GVT, which harbors the amino acids 1-123 of GM-CSF whose L114L115V116 was substituted with G114V115T116, was evidently higher than that of the DT386-GMCSF, whereas the specific cytotoxicity to blast recovered from mice injected with HL60, a cell line highly expresses the GM-CSFR, was similar. These results have provided an important basis for the future development of the immunotoxins targeting the GM-CSFR.
%K Immunotoxin Diphtheria toxin Granulocyte-macrophage colony-stimulating factor Prokaryotic expression Mutant<br>免疫毒素
%K 白喉毒素
%K 粒细胞-巨噬细胞集落刺激因子
%K 原核表达
%K 突变体
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=951380788B355DEA7D75520FA3B199C9&aid=38175C0F6B82002D&yid=A732AF04DDA03BB3&vid=DB817633AA4F79B9&iid=B31275AF3241DB2D&sid=16D8618C6164A3ED&eid=94E7F66E6C42FA23&journal_id=1671-8135&journal_name=中国生物工程杂志&referenced_num=0&reference_num=21