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当前位置: 首页 > 文章 > 鸡白介素18与鸡γ干扰素基因融合表达载体构建 黑龙江八一农垦大学学报 2017,29 (1) 49-53+82
Position: Home > Articles > Construction of Chicken Interleukin 18 and Interferon Gamma Gene Fused Expression Vector Journal of Heilongjiang Bayi Agricultural University 2017,29 (1) 49-53+82

鸡白介素18与鸡γ干扰素基因融合表达载体构建

作  者：
李晓婷;宋佰芬;崔玉东
单  位：
黑龙江八一农垦大学动物科技学院;黑龙江八一农垦大学生命科学技术学院
关键词：
鸡白细胞介素18;鸡干扰素γ;overlap;原核表达载体
摘  要：
为了研究新型细胞因子制剂,构建鸡白细胞介素18和鸡干扰素γ的基因融合表达载体。无菌提取鸡脾细胞的总RNA,以其为模板经过RT-PCR的方法分别扩增出Ch IL-18和Ch IFN-γ基因片段,并连接到p MDTM18-T克隆载体上,构建重组质粒p MDTM18-T-Ch IL-18和p MDTM18-T-Ch IFN-γ。以重组质粒p MDTM18-T-Ch IL-18和p MDTM18-T-Ch IFN-γ为模板,利用多肽氨基酸接头和overlap技术,获得Ch IL-18-Ch IFN-γ融合基因并连接到克隆载体p MDTM18-T上,经PCR、酶切及测序鉴定正确后连接到p ET32a原核表达载体中。构建Ch IL-18-Ch IFN-γ原核表达载体。为下一步研究二者之间的作用关系及抗病毒功能奠定了基础。
译  名：
Construction of Chicken Interleukin 18 and Interferon Gamma Gene Fused Expression Vector
作  者：
Li Xiaoting;Song Baifen;Cui Yudong;College of Animal Science and Technology,Heilongjiang Bayi Agricultural University;College of Life Science and Technology,Heilongjiang Bayi Agricultural University;
关键词：
Chicken Interleukin 18;;chicken interferon gamma;;overlap;;fused expression vector
摘  要：
In order to study the new cytokine preparation,the fused expression vectors of chicken interleukin 18 and interferon-γgene were constructed. Total RNA of chicken spleen cells was extracted aseptically. Ch IL-18 and Ch IFN-γ gene fragments were amplified by RT-PCR method,then cloned into p MDTM18-T and constructed recombinant plasmid p MDTM18-T-Ch IL-18 and p MDTM18-T-Ch IFN-γ. Fused gene of Ch IL-18-Ch IFN-γ was obtained by peptide amino acid linker,overlap technology was used to recombinant plasmid p MDTM18-T-Ch IL-18 and p MDTM18-T-Ch IFN-γ as template.,Ch IL-18-Ch IFN-γ were cloned into p MDTM18-T vector and were identified by PCR,digestion and sequencing. Positive plasmid was digested by restriction endonuclease and fused gene was inserted into p ET32 aprokaryotic expression vector.The prokaryotic expression vector of Ch IL-18-Ch IFN-γ was successfully constructed. It laid the foundation for studying the role of their relationship and the antiviral function.

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