| 狂犬病病毒3aG株核蛋白基因cDNA的克隆、序列分析及在COS细胞中的暂态表达 |
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| 引用本文： | 李伟,张新梅,张曼夫,王树惠.狂犬病病毒3aG株核蛋白基因cDNA的克隆、序列分析及在COS细胞中的暂态表达[J].中国兽医学报,1999(4). |
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| 作者姓名： | 李伟 张新梅 张曼夫 王树惠 |
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| 作者单位： | 中国农业大学生物学院!北京100094 |
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| 摘 要： | 从狂犬病病毒 ３ａ Ｇ 株感染的 Ｂ Ｈ Ｋ 细胞裂解上清液中快速提取病毒 Ｒ Ｎ Ａ，用 Ｒ Ｔ Ｐ Ｃ Ｒ 得到编码核蛋白完整结构基因的 ｃ Ｄ Ｎ Ａ。进一步将此基因克隆入 ｐ Ｕ Ｃ １８中，进行核苷酸序列分析，并与国外狂犬病病毒 Ｐ Ｖ、 Ｃ Ｖ Ｓ、 Ｓ Ａ Ｄ Ｂ１ ９ 株以及国内另一狂犬病病毒 ５ａ Ｇ 株的 Ｎ 基因序列进行了同源性比较。然后将 Ｎ 基因正向插入真核表达质粒中，转染 Ｃ Ｏ Ｓ７ 细胞，用免疫组化方法证明所克隆基因可在细胞中正确表达出 Ｎ 蛋白。
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| 关 键 词： | 狂犬病病毒 核蛋白基因 RT-PCR cDNA 真核表达 免疫组化 |
Cloning,Sequencing of the Rabies Virus Nucleoprotein Structural Gene and Its Transient Expression in Cos 7 Cells |
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| Li Wei Zhang Xinmei,Zhang Manfu Wang Shuhui.Cloning,Sequencing of the Rabies Virus Nucleoprotein Structural Gene and Its Transient Expression in Cos 7 Cells[J].Chinese Journal of Veterinary Science,1999(4). |
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| Authors: | Li Wei Zhang Xinmei Zhang Manfu Wang Shuhui |
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| Abstract: | The nucleoprotein structural gene cDNA of rabies virus 3aG strain was amplified by RT PCR from genomic RNA and cloned into pUC18 plasmid vector.Its nucleotide sequence was analyzed.The nucleotide sequences homology between 3aG strain and PV,CVS,SADB19 and 5aG strain were determined to be 92.46%, 92.17%, 92.69% and 94.23% respectively.The cloned cDNA was further inserted into eukaryotic expression plasmid vector pcDNA3.1( ) and transfected into COS 7 cells.The expression of N protein was then detected by immunostaining and showed to be positive. |
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| Keywords: | rabies virus nucleoprotein structural gene RT PCR cDNA eukaryotic expression immunostaining |
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