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2025年7月29日 星期二
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粒细胞-巨噬细胞集落刺激因子与生长抑素融合表达质粒(pGM-CSF/SS)的构建及其对小鼠的免疫
舒邓群 茆达干 杨利国 曹少先 吴志敏
江西农业大学动物科技学院 南京农业大学动物科技学院 华中农业大学动物科技学院 南京农业大学动物科技学院 江西农业大学动物科技学院
Construction and Immunization of Fusion-expression Plasmid Harboring Porcine GM-CSF and Somatostatin
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摘要 从猪真核表达质粒pGM-CSF通过PCR扩增出猪GM-CSF基因的编码序列,将GM-CSF基因亚克隆到生长抑素真核表达质粒pcS/2SS中,构建GM-CSF与生长抑素(SS)的融合表达质粒pGM-CSF/SS,酶切鉴定和DNA测序证实重组的融合表达质粒pGM-CSF/SS构建正确。选择30只小鼠,随机分为三组,其中1组为对照组,2组和3组分别肌肉注射pcS/2SS和pGM-CSF/SS,研究pGM-CSF/SS对小鼠的免疫效果。结果表明:GM-CSF可增强小鼠脾脏淋巴细胞的增殖能力,以pGM-CSF/SS组增殖能力最强,分别比对照组和pcS/2SS提高8.4%(P<0.05)。肌肉注射pcS/2SS和pGM-CSF/SS免疫小鼠均能有效激发免疫系统,产生较强的免疫应答,与对照组相比,生长抑素抗体P/N值都有所提高,且随着免疫时间的延长,呈现不断增高的趋势;pGM-CSF/SS组P/N在试验的各个时期均高于pcS/2SS组。阳性鼠的比例以肌注pGM-CSF/SS为最高,达60%,pcS/2SS肌注组为30%,对照组没有出现阳性鼠;这些结果证明,GM-CSF对生长抑素DNA疫苗有一定的免疫增强作用,为研究GM-CSF基因对生长抑素DNA疫苗的免疫增强作用提供了理论基础。
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舒邓群茆达干杨利国曹少先吴志敏
关键词 粒细胞-巨噬细胞集落刺激因子生长抑素融合表达质粒免疫    
Abstract:Porcine GM-CSF gene was amplified from porcine expression plasmid pGM-CSF by PCR. Fragment containing GM-CSF gene was subcloned into somatostatin eukaryotic expression plasmid pcS/2SS to construct fusion-expression plasmid (pGM-CSF/SS) harboring porcine GM-CSF and somatostatin (SS). Restriction endonucleases digestion and DNA sequencing showed that the fusion-expression plasmid (pGM-CSF/SS) harboring porcine GM-CSF and somatostatin was successfully constructed. To study the immunization effect of pGM-CSF/SS on mice, a total of 30 mice at the age of 22 days were divided into 3 groups and immunized with salt solution (group 1), pcS/2SS (group 2) and pGM-CSF/SS (group 3), respectively. The results showed that lymphocyte proliferation response of pGM-CSF/SS was higher than that of the control group and pcS/2SS alone (P<0.05), indicating that GM-CSF could enhance lymphocyte proliferation responses. Fusion-expression plasmid of GM-CSF with SS significantly enhanced the P/N value of SS antibody compared to the control group and immunization of pcS/2SS alone. Percentage of mice with positive SS antibody immunized with pGM-CSF/SS was higher than that of other groups, which got to 60%. Percentage of that immunized with pcS/2SS was 30%. In conclusions, genetic adjuvant GM-CSF can enhance immune response of somatostatin DNA vaccine, which is an access to further research for immuno-enhancement of GM-CSF gene on somatostatin DNA vaccine.
Key wordsGranulocyte-macrophage colony-stimulating factor(GM-CSF)    Somatostatin (SS)    Fusion-expression plasmid    Immunization
收稿日期: 2007-05-14     
通讯作者: 舒邓群   
引用本文:   
舒邓群 茆达干 杨利国 曹少先 吴志敏. 粒细胞-巨噬细胞集落刺激因子与生长抑素融合表达质粒(pGM-CSF/SS)的构建及其对小鼠的免疫[J]. , 2008, 16(1): 0-.
. Construction and Immunization of Fusion-expression Plasmid Harboring Porcine GM-CSF and Somatostatin. , 2008, 16(1): 0-.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/      或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2008/V16/I1/0
 
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