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| 猪白细胞介素-4基因的克隆与表达 | |
| 引用本文： | 陈国华,景志忠,孙世铎,罗启慧,窦永喜,蒙学莲.猪白细胞介素-4基因的克隆与表达[J].中国预防兽医学报,2005,27(6):469-473. |
| 作者姓名： | 陈国华 景志忠 孙世铎 罗启慧 窦永喜 蒙学莲 |
| 作者单位： | 1. 中国农业科学院兰州兽医研究所,甘肃,兰州,730046 2. 西北农林科技大学动物科技学院,陕西,杨凌,712100 3. 中国农业科学院兰州兽医研究所,甘肃,兰州,730046;西北农林科技大学动物科技学院,陕西,杨凌,712100 |
| 基金项目： | 新材料领域项目，引进国际先进农业科技计划(948计划) |
| 摘 要： | 利用RT-PCR技术,从被刺激诱导的PBMC中克隆IL-4基因,序列分析表明:克隆的猪IL-4基因序列与GenBank上登录的IL-4基因的核苷酸和氨基酸序列同源性分别为99%和97.8%.然后用表达型引物从T载体上扩增IL-4基因,双酶切PCR产物后与表达载体pGEX连接,构建重组表达质粒pGEX-IL-4,用IPTG诱导表达,表达产物经SDS-PAGE分析表明,表达出38 ku融合蛋白,占菌体总蛋白的30%以上,并且以包涵体的形式存在,这为猪重组IL-4规模化生产和疫苗佐剂的研制奠定了基础. |
| 关 键 词： | 猪IL-4基因 克隆 表达 |
| 文章编号： | 1008-0589(2005)06-0469-05 |
| 收稿时间： | 2004-08-16 |
| 修稿时间： | 2004年8月16日 |
Cloning and expression of porcine IL-4 gene |
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| CHEN Guo-hua,JING Zhi-zhong,SUN Shi-duo,LUO Qi-hui,DOU Yong-xi,MENG Xue-lian.Cloning and expression of porcine IL-4 gene[J].Chinese Journal of Preventive Veterinary Medicine,2005,27(6):469-473. | |
| Authors: | CHEN Guo-hua JING Zhi-zhong SUN Shi-duo LUO Qi-hui DOU Yong-xi MENG Xue-lian |
| Institution: | 1.Lanzhou Veterinary Research Institute, Chinese Academy of Aagricukural Sciences, Lanzhou 730046, China; 2.College of Animal Science and Technology,Northwest Science Techology University of Angriculture and Forestry,YangLing 712100,China |
| Abstract: | Using RT-PCR,pig IL-4 gene was cloned from porcine PBMC stimulated by LPS and PHA,After sequencing,compared with IL-4 gene published in GenBank,the homologies of the nucleotide acide sequence and amino acide sequence were 99?% and(97.8?%),respectively.Then the recombinant expression plasmid(pGEX-IL-4) was constructed,pGEX-Il-4 was expressed in E.coli by IPTG induction,SDS-PAGE analysis showed that the expression product was fusion protein with molecular weight of 38 ku and the percentage of expression protein in E.coil protein was 30?%.This has laid a foundation for the production and vaccine adjuvant development of porcine IL-4. |
| Keywords: | porcine interleukin-4(IL-4)gene cloning expression |
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