| 新型鸡白细胞介素15真核表达质粒构建及其体外表达 |
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| 引用本文： | 马德星,李广兴,马春丽,杨静红.新型鸡白细胞介素15真核表达质粒构建及其体外表达[J].中国家禽,2011,33(18). |
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| 作者姓名： | 马德星 李广兴 马春丽 杨静红 |
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| 作者单位： | 1. 东北农业大学动物医学学院,黑龙江哈尔滨,150030
2. 东北农业大学食品学院,黑龙江哈尔滨,150030 |
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| 基金项目： | 国家自然科学青年基金项目(30901061); 东北农业大学科学研究基金(2010RCB38) |
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| 摘 要： | 应用重叠扩增PCR(SOE PCR)技术将鸡白细胞介素15(ChIL-15)的冗长信号肽序列替换为较短的鸡白细胞介素2(ChIL-2)信号肽序列,构建了一种新型ChIL-15融合基因(NChIL-15).将其连接到克隆载体pMD18-T中得到重组克隆质粒pMD18T-NChIL-15.阳性克隆质粒经鉴定正确后将NChIL-15基因亚克隆入真核表达载体pcDNA3.1(+)中,得到重组阳性表达质粒pcDNA3.1(+)-NChIL- 15.阳性表达质粒经鉴定正确后应用磷酸钙法体外转染293T细胞,通过间接免疫荧光试验(IFA)检测到了NChIL-15蛋白在293T细胞中的表达.本研究为NChIL-15佐剂作用的深入研究奠定了基础.
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| 关 键 词： | NChIL-15 转染 间接免疫荧光试验 |
Construction of Eukaryotic Expressing Plasmid of New Chicken Interleukin 15 and Its Expression in vitro |
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| MA Dexing,LI Guangxing,MA Chunli,YANG Jinghong.Construction of Eukaryotic Expressing Plasmid of New Chicken Interleukin 15 and Its Expression in vitro[J].China Poultry,2011,33(18). |
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| Authors: | MA Dexing LI Guangxing MA Chunli YANG Jinghong |
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| Institution: | MA Dexing1,LI Guangxing1,MA Chunli2,YANG Jinghong1(1.College of Veterinary Medicine,Northeast Agricultural University,Harbin,Heilongjiang 150030,2.Food College,Heilongjiang 150030) |
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| Abstract: | A new type gene of chicken interleukin 15(NChIL-15) was amplified by gene splicing overlap extension PCR(SOE PCR).NChIL-15 was cloned into cloning vector pMD18-T,and the recombinant cloning plasmid was sequenced and named pMD18T-NChIL-15.NChIL-15 gene was then sequenced and cloned into eukaryotic expressing vector pcDNA3.1(+).The recombinant expressing plasmid pcDNA3.1(+)-NChIL-15 was identified and then transfected into 293T cell in vitro with Ca3(PO4)2.The expression of NChIL-15 protein was successfully d... |
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| Keywords: | NChIL-15 transfected indirect immun of fluorescence assay |
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