| 陕西烟区烟草脉带花叶病毒外壳蛋白基因的克隆和原核表达 |
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| 引用本文： | 刘萍,纪玲玲,郝兴安,吴云锋,成巨龙.陕西烟区烟草脉带花叶病毒外壳蛋白基因的克隆和原核表达[J].中国烟草学报,2011,17(5):87-89,95. |
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| 作者姓名： | 刘萍 纪玲玲 郝兴安 吴云锋 成巨龙 |
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| 作者单位： | 1. 西北农林科技大学植保学院与旱区作物逆境生物学国家重点实验室,杨凌,712100
2. 陕西省烟草研究所,西安,710061 |
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| 基金项目： | 中国烟草总公司科技重点项目，陕西省烟草公司科学研究与技术开发项目，高等学校学科创新引智计划 |
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| 摘 要： | 根据烟草脉带花叶病毒(TVBMV)CP序列合成上下游引物,利用RT-PCR方法获得了(TVBMV)CP基因。序列分析表明,TVBMV CP基因全长813nt,编码271个氨基酸;与GenBank中已报道的9个分离CP基因相比,其核苷酸及氨基酸序列同源性分别为90.28%-98.15%和97.05%-100%。将TVBMV CP基因经BamHI/NotI双酶切定向插入到pET30a载体中,构建了原核表达载体pET30-TVBMV CP,重组质粒经BamHI和NotI双酶切鉴定及基因测序验证正确后,用IPTG进行诱导表达。结果表明:TVBMV CP在大肠杆菌中获得了高效表达,表达产物的分子量为37 kD。用表达产物为抗原免疫家兔制备了特异性抗血清,其效价为1/2048。用Western-blot检测田间样品,其结果表明,制备的抗血清可用于检测田间的发病植株。
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| 关 键 词： | 烟草脉带花叶病毒 外壳蛋白 原核表达 抗血清 |
Cloning and prokaryotic expression of the coat protein gene of tobacco vein banding mosaic virus from Shaanxi province |
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| LIU Ping,JI Ling-ling,HAO Xing-an,WU Yun-feng,CHENG Ju-long.Cloning and prokaryotic expression of the coat protein gene of tobacco vein banding mosaic virus from Shaanxi province[J].Acta Tabacaria Sinica,2011,17(5):87-89,95. |
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| Authors: | LIU Ping JI Ling-ling HAO Xing-an WU Yun-feng CHENG Ju-long |
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| Affiliation: | LIU Ping1,JI Ling-ling1,HAO Xing-an1,WU Yun-feng1,CHENG Ju-long21 College of Plant Protection and State Key Laboratory of Crop Stress Biology in Arid Areas,Northwest A&F University,Yangling 712100,China,2 Shaanxi Tobacco Institute,Xi'an 710061 |
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| Abstract: | The coat protein(CP) gene of Tobacco Vein Banding Mosaic Virus(TVBMV) from Shaanxi province was obtained by RT-PCR.Sequence analysis showed that TVBMV CP gene was 813 nt long and encoded a 271 amino acid protein.Homology analyzed between TVBMV CP and other 9 strains in GenBank were 90.28%-98.15% at nucleotide level,and 97.05%-100% at amino acid level.The CP gene was cloned into pMD18-T Simple Vector and inserted into pET30a which were cleaved with restrict endonuclease BamH I/Not I.The expression vector of ... |
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| Keywords: | tobacco vein banding mosaic virus coat protein prokaryotic expression antiserum |
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