| PRRSV陕西分离株ORF3基因的克隆、序列分析及原核表达载体的构建 |
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| 引用本文： | 张 琦,王 菡,黎 径,梁 靓,许信刚.PRRSV陕西分离株ORF3基因的克隆、序列分析及原核表达载体的构建[J].西北农业学报,2010,19(10):21-26. |
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| 作者姓名： | 张 琦 王 菡 黎 径 梁 靓 许信刚 |
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| 作者单位： | 1. 陕西出入境检验检疫局检验检疫技术中心,西安,710068
2. 西北农林科技大学,动物医学院,陕西杨凌,712100 |
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| 基金项目： | 陕西省自然科学基金项目(2006C111); 陕西省科技攻关项目(2009K02-01) |
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| 摘 要： | 根据GenBank公布的PRRSVORF3基因的核苷酸序列,设计并合成一对特异性引物,用RT-PCR方法扩增PRRSV陕西分离株ORF3基因,将其克隆入pGEM-T载体中,测序并进行序列分析。再将ORF3基因亚克隆入pET-32a中,构建原核表达载体。结果扩增到765 bp的PRRSV全长ORF3基因,序列分析结果表明,分离株与SY0608亲缘关系较近,而与CH-2、HB-2关系较远。重组原核表达质粒经酶切鉴定正确后命名为pET-GP3,为进一步研究GP3蛋白的原核表达、免疫特性、结构与功能奠定了基础。
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| 关 键 词： | 猪繁殖与呼吸综合征病毒 陕西分离株 ORF3基因 序列分析 原核表达载体 |
Cloning, Sequence Analysis and Construction of Prokaryotic Expression Vector of ORF3 Gene of PRRSV Shaanxi Strain |
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| ZHANG Qi,WANG Han,LI Jing,LIANG Liang and XU Xingang.Cloning, Sequence Analysis and Construction of Prokaryotic Expression Vector of ORF3 Gene of PRRSV Shaanxi Strain[J].Acta Agriculturae Boreali-occidentalis Sinica,2010,19(10):21-26. |
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| Authors: | ZHANG Qi WANG Han LI Jing LIANG Liang and XU Xingang |
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| Institution: | ZHANG Qi1,WANG Han1,LI Jing1,LIANG Liang1 and XU Xingang2 (1.Inspection and Quarantine Technical Centre,Shaanxi Entry-Exit Inspection and Quarantine Bureau of the People's Republic of China,Xi'an 710068,China,2.College of Veterinary Medicine,Northwest A&F University,Yangling Shaanxi 712100,China) |
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| Abstract: | According to the published complete nucleotide sequence of PRRSV(Porcine reproductive and respiratory syndrome virus) in GenBank,a pair of primers specific to the full-length of ORF3 gene encoding the envelope glycoprotein GP3 were designed.The ORF3 gene fragment amplified by RT-PCR was cloned into pGEM-T vector.The recombinant plasmid was sequenced and ORF3 gene was compared with GenBank's other PRRSV strains.Then the gene was cloned into vector pET-32a for prokaryotic expression in E.coli.The result showe... |
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| Keywords: | Porcine reproductive and respiratory syndrome virus(PRRSV) Shaanxi strain ORF3 Sequence analysis Prokaryotic expression |
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