检索结果-南通市图书馆

Expression of Human Vascular Endothelial Growth Factor Receptor Flt-1 Extracellular Domain 1-3 Loop cDNA in Pichia pastoris, Purification of the expressed product and Detection of Its Biological Activity

引用

Biomedical and Environmental Sciences 2001年第4期14卷 292-301页

作者： MALI ZHANGZHI－QING 等 InstituteofMolecularImmunology ThefirstMilitaryMedicalUniversityGuangzhou510515China StateKeyLaboratoryofMolecularVirologyandGeneticEngineering Beijing100052China

Objective To express human vascular endothelial growth factor receptor Flt-1 extracellular domain 1-3 loop cDNA in Pichia. pastroris, and to purify the expressed product and detect its biological activity. Methods By inserting human Flt-1 (1-3 loop) cDNA coding 316 amino acid residues into Pichia pastoris expression vector pPIC9K containing AOX1 promoter and the sequences of α secreting signal peptides, a recombinant expression plasmid pPIC9K/Flt-1 (1-3) was constructed and transformed to yeast host strain GS115, then His + Mut s phenotype transformant was screened out and cultured in flasks, and Flt-1 (1-3) was expressed under the induction of 1% methanol. Results SDS-PAGE showed that after being induced with 1% methanol for 4d, the expressed product existed in supernatant in the form of soluble molecule and contained 60% of total protein expressed. Western blot showed good antigenicity and specificity of expressed product. After being purified by CM-Sepharose FF and Sephacryl S-100 chromatography, the purity of the expressed product reached above 90%. Biological assay proved that the expressed product could bind to hVEGF 165 and inhibit the proliferation of HUVEC stimulated by hVEGF 165. Conclusion Human vascular endothelial growth factor receptor Flt-1 extracellular domain 1-3 loop was successfully expressed. The study lays a foundation for further application of the expressed product in the treatment of vasoformation related diseases, such as tumor and diabetic retinopathy.

关键词： Endothelial Growth Factor Pichia pastoris expressed product growth factor receptor diabetic retinopathy biological activity amino acid residues expression vector product in form of

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Expression of Human Vascular Endothelial Growth Factor (VEGF_(165)) in Pichia pastoris and Its Biological Activity

引用

Biomedical and Environmental Sciences 2001年第4期14卷 302-311页

作者： MALI ZHANGZHI－QING 等 InstituteofMolecularImmunology theFirstMilitaryMedicalUniversityGuangzhou510515China StateKeyLaboratoryofMolecularVirologyandGeneticEnginering Beijing100052China

Objective To express human vascular endothelial growth factor (hVEGF-{165}) cDNA in %Pichia pastroris, %purify the expressed product and detect the biological activity of it. Methods By inserting hVEGF-{165} cDNA coding 165 amino acid residues into %Pichia pastoris% expression vector pPIC9K containing AOX1 promoter and the sequences of α secreting signal peptides, a recombinant expression plasmid pPIC9K/hVEGF-{165} was constructed and transformed to yeast host strain KM71, then multiple_copy insert transformants were screened out and cultured in flasks, and hVEGF-{165} was expressed under the induction of 1% methanol. Results SDS_PAGE showed that after being induced with 1% methanol for 4d, the expressed product existed in supernatant in the form of soluble molecule and contained 60% of total protein expressed. Western blot showed good antigenicity and specificity of expressed product. After being purified by Heparin_Sepharose CL6B affinity chromatography, the purity of expressed product reached above 90%. Biological assays proved that the expressed product could stimulate the proliferation of HUVEC. Conclusion hVEGF-{165} was successfully expressed. The study opened up a wide prospect for the application of VEGF-{165} in the prevention and treatment of ischemic heart disease and other tissue ischemic diseases such as secondary arterial occlusion in limbs.

关键词： Endothelial Growth Factor Biological Activity expressed product endothelial growth factor prevention and treatment affinity chromatography ischemic heart disease biological activity amino acid residues expression vector Pichia pastoris form of up a

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Immune responses to the expressed products of the CSP antigen gene of Plasmondium falciparum south in China isolate FCC1/HN in Hela cell

引用

南通大学学报(医学版) 2003年第1期 115-115页

作者： Yu XB Liu YW Luo SH

Objective:To construct a eukaryotic expression system with pcDNA3-PfCSP/Hela for the Circumsporozoite protein(CSP) gene of Plasmodium falciparum(***),to observe the immune responses in BALB/c mice induced by the expressed proteins. Methods:The recombinant plasmid pcDNA3-PfCSP was transformed into the Hela cell *** expressed protein was isolated and analyzed by using SIDS-PAGE and used for immunization of BALB/c mice by subcutaneous, intravenous, and intraperitoneal ***-linked immunosorbent assay(ELISA),Dot-ELISA,Western blot,T lymphocyte proliferation test,natural killer cell(NKC) activity assay,and CD4(+) and CD8(+) T cell detection were used for observation of humoral and cellular immune responses. Results:Immune sera strongly reacted with the expressed protein,antibody titer was up to 1:6400 as detected by *** blot analysis revealed a specific band at 38.3 *** the spleen cells of normal and immunized BALB/c mice were specifically stimulated with expressed protein,the optical densities were 0.12+/-0.03 and 0.34+/-0.04,*** latter were significantly higher than the former(Pproducts made with a eukaryotic expression sytem could be specifically detected in BALB/c *** findings indicated that the expressed protein could enhance the immune function in mice.

关键词： plasmodium falciparum circumsporozoite protein Held cell expressed product immune response

来源：

同方期刊数据库评论

在线全文

同方期刊数据库

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：