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| 鸡Mx蛋白基因诱变修饰及抗病活性 | |
| 引用本文： | 倪黎纲,吴晓伟,程旭梅,陈昊,陈强,宋成义,徐琪,李碧春.鸡Mx蛋白基因诱变修饰及抗病活性[J].微生物学报,2008,48(6):785-789. |
| 作者姓名： | 倪黎纲 吴晓伟 程旭梅 陈昊 陈强 宋成义 徐琪 李碧春 |
| 作者单位： | 1. 扬州大学动物科学与技术学院,扬州,225009 2. 苏州大学医学院基础医学系,苏州,215006 3. 扬州大学动物科学与技术学院,扬州,225009;江苏省农业科学院畜牧研究所,南京,210014 |
| 基金项目： | 国家自然科学基金 , 教育部高等学校博士学科点专项科研基金 |
| 摘 要： | 目的]进一步研究鸡Mx蛋白第631位氨基酸的变异与鸡群抗病性的相关性.方法]本实验利用PCR突变技术将鸡Mx蛋白基因的全长cDNA第2032位的碱基由G突变为A(既631位氨基酸的改变),并将突变的Mx基因插入真核表达载体pcDNA3.0,重组表达载体转染COS-Ⅰ细胞后,进行RT-PCR与间接免疫荧光(IFA)鉴定.结果]对鸡Mx蛋白基因的cDNA进行PCR诱变修饰正确,构建了能够正确表达鸡Mx蛋白的重组真核表达载体;诱变修饰重组Mx蛋白对抗新城疫病毒(NDV)感染分析结果显示,重组Mx蛋白具有较强的抗新城疫病毒生物活性.结论]为下一步研究鸡Mx蛋白的抗病机理与制备抗病转基因鸡奠定了坚实的基础. |
| 关 键 词： | Mx蛋白 PCR突变 抗病毒活性 新城疫病毒 蛋白基因 基因诱变 修饰 抗病活性 specificity antiviral Identification chicken Gene modified 抗病转基因 抗病机理 生物活性 显示 感染分析 病毒 新城疫 重组真核表达载体 正确表达 结果 |
| 文章编号： | 0001-6209(2008)06-0785-05 |
| 收稿时间： | 2007/11/29 0:00:00 |
| 修稿时间： | 2007年11月29 |
Mutagenesis modified of Mx Gene from chicken and Identification of its antiviral specificity |
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| Ligang Ni,Xiaowei Wu,XumeiCheng,Hao Chen,Qiang Chen,Chengyi Song,Qi Xu and Bichun Li.Mutagenesis modified of Mx Gene from chicken and Identification of its antiviral specificity[J].Acta Microbiologica Sinica,2008,48(6):785-789. | |
| Authors: | Ligang Ni Xiaowei Wu XumeiCheng Hao Chen Qiang Chen Chengyi Song Qi Xu and Bichun Li |
| Institution: | College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;Medical School of Suzhou University, Suzhou University, Suzhou 215000, China;College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;(1College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;3Institute of Animal Husbandry and Veterinary, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China) |
| Abstract: | OBJECTIVE: To study whether the antiviral specificity of chicken Mx protein is determined by an amino acid substitution at position 631. METHODS: We used PCR site-directed mutagenesis technique by which a single amino acid was reciprocally substituted G with A at position 2032bp of chicken Mx cDNA. Sequence analysis confirmed successful mutation from G to A at 2032bp of chicken Mx cDNA. The fragments amplified by PCR containing the mutation site were subcloned into a enteukaryotic expression vector. Then the recombinant vector was transfected into COS-I cell, Mx gene and Mx protein in the transfected COS-I cell were detected by RT-PCR and indirect fluorescence assay. RESULTS: The results showed that COS- I cell transfected the recombinant plasmid could stably express the Mx protein. The antivirus assay showed that Mx protein had characteristics of resistance to infection of Newcastle Disease Virus. CONCLUSION: This study may provide a basis of the virus-resistant mechanism of Mx protein and production of virus-resistant transgenic chicken. |
| Keywords: | Mx protein PCR Mutagenesis Virus-resistant activity Newcastle Disease Virus |
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