| 卵泡抑制素与GFP融合基因疫苗pEGISI的构建及表达 |
| |
| 引用本文： | 曹少先,茆达干,舒邓群,程宝,王进荣,刘铁铮,杨利国.卵泡抑制素与GFP融合基因疫苗pEGISI的构建及表达[J].家畜生态,2007(2). |
| |
| 作者姓名： | 曹少先 茆达干 舒邓群 程宝 王进荣 刘铁铮 杨利国 |
| |
| 作者单位： | 江苏省农业科学院畜牧研究所，南京农业大学动物科技学院，南京农业大学动物科技学院，南京农业大学动物科技学院，南京农业大学动物科技学院，江苏省农业科学院畜牧研究所，南京农业大学动物科技学院 江苏南京210014，江苏南京210095，江苏南京210095，江苏南京210095，江苏南京210095，江苏南京210014，江苏南京210095，华中农业大学动物科技学院，湖北武汉430070 |
| |
| 基金项目： | 国家自然科学基金资助项目(30270959)，高校博士点基金资助项目(2000030716) |
| |
| 摘 要： | 为构建卵泡抑制素与绿色荧光蛋白(GFP)融合基因疫苗pEGISI,将pcISI中的乙肝表面抗原(HBsAg)S基因及插入S基因中的抑制素(INH)基因片段酶切回收;PCR扩增pcISI中INH基因片段,调整阅读框,一起融合到pEGFP-N1中EGFP基因的5'端,构建ISI-EGFP融合表达质粒pEGISI。酶切和测序鉴定表明,重组质粒pEGISI构建成功。将pEGISI转染293T细胞,16h后检测到绿色荧光,48h检测到强烈荧光,说明融合基因在293T细胞获得高表达;ELISA检测证实,表达产物ISI-EGFP融合蛋白具有INH的抗原抗体反应原性。质粒pEGISI的成功构建及表达为抑制素基因免疫的机理及安全性研究奠定了基础。
|
| 关 键 词： | 卵泡抑制素 EGFP 基因克隆 基因表达 |
Construction and Expression of Inhibin DNA Vaccine pEGISI in 293T Cell |
| |
| CAO Shao-Xian,MAO Da-gan,SHU deng-qun,CHENG Bao,WANG Jin-Rong,LIU Tie-zheng,YANG Li-Guo.Construction and Expression of Inhibin DNA Vaccine pEGISI in 293T Cell[J].Ecology of Domestic Animal,2007(2). |
| |
| Authors: | CAO Shao-Xian MAO Da-gan SHU deng-qun CHENG Bao WANG Jin-Rong LIU Tie-zheng YANG Li-Guo |
| |
| Institution: | CAO Shao-Xian1,MAO Da-gan2,SHU deng-qun2,CHENG Bao2,WANG Jin-Rong2,LIU Tie-zheng1,YANG Li-Guo2,3 |
| |
| Abstract: | To construct inhibin(INH) expression vector fused GFP,HBsAg S gene incorporated INH fragment was collected from pcISI by restriction endonuclease digestion,another copy of INH fragment was amplified by PCR to adjust the open-reading frame of EGFP.Then those fragments were ligated and cloned into the 5' end of EGFP gene of pEGFP-N1,and ISI-EGFP recombinant plasmid is pEGISI.This plasmid was identified by restriction endonuclease digestion and sequencing,then the pEGISI was transfected into 293T cells.The recombinant protein was detected by ELISA and fluorescence microscope.Green fluorescence was observed at 16h after transfection and became stronger at 48h.It was suggested that the recombinant gene expressed in 293T cells.The fused protein reacted with the antibodies against INH.In conclusion,successfully construction and expression of the plasmid pEGISI will provid the basis for the study of immunity mechanism and safety of somatostatin gene vaccine. |
| |
| Keywords: | Inhibin EGFP gene clone gene expression |
| 本文献已被 CNKI 等数据库收录！ |
|
