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当前位置: 首页 > 文章 > 新兴猪γ-干扰素基因克隆及其真核表达质粒的构建 动物医学进展 2005,26 (9) 78-81
Position: Home > Articles > Clone of Xinxing Porcine Interferon-γ Gene and Construction of Its Eukaryotic Expression Plasmids Progress in Veterinary Medicine 2005,26 (9) 78-81

新兴猪γ-干扰素基因克隆及其真核表达质粒的构建

作  者：
温纳相;陈瑞爱;裴仉福;唐满华;朱文冠;程含波;李琳
单  位：
广东新华农动物保健品有限公司
关键词：
猪;γ-干扰素;克隆;真核表达质粒
摘  要：
为研发猪γ-干扰素(interferon-gam-ma,IFN-γ)制剂,根据已发表的猪IFN-γ基因序列设计一对引物,应用RT-PCR技术从3周龄新兴猪脾细胞中扩增出约500 bp的基因片段;将其片段连接到pMD18-T载体,经PCR、酶切及序列测定表明,该基因片段由501个核苷酸组成,共编码166个氨基酸,与NCBI GenBank上登载的猪IFN-γ基因序列完全一致。新兴猪IFN-γ基因的成功克隆及真核表达质粒的构建,将为进一步研发猪干扰素制剂奠定了基础。
译  名：
Clone of Xinxing Porcine Interferon-γ Gene and Construction of Its Eukaryotic Expression Plasmids
作  者：
WEN Na-xiang,CHEN Rui-ai,PEI Zhang-fu,TANG Man-hua,ZHU Wen-guan,CHENG Han-bo,LI Lin(Guangdong New Huanong Animal Health Products Corporation Limited,Xinxing,Guangdong,527439,China)
关键词：
pig;IFN-γ;clone;eukaryotic expression plasmid
摘  要：
In this study,one specific pair of primers was designed and synthesized according to the sequence of porcine IFN-γ gene published NCBI GenBank.The total RNA was extracted from 3-week-old porcine spleen.Porcine IFN-γ was amplified by reverse transcription-polymerase chain reaction(RTPCR).The gene was inserted in pMD-18 T vector and the recombinant plasmids were transformed into the E.Coli DH5α.The positive clones were selected and incubated.After they were identified by restriction endonuclease analysis,PCR and sequence analysis.The result indicated that the full-length of porcine IFNγ cDNA was 501 bp,encoding 166 amino acids.The sequence was just the same as the published one from NCBI GenBank.The clone of porcine IFN-γ and the construction of eukaryotic expression plasmid pcDNA3.1-IFNγ have laid a foundation for the production and development of porcine IFN-γ.

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