| 小反刍兽疫病毒疫苗株N和M基因的克隆、序列分析及原核表达 |
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| 引用本文：何亚鹏，鲍长磊，张 琪，付明哲，许信刚.小反刍兽疫病毒疫苗株N和M基因的克隆、序列分析及原核表达[J].西北农业学报,2017,(2):179~184 |
| DOI:10.7606/j.issn.1004-1389.2017.02.004 |
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| 基金项目:陕西省科技攻关（2015NY162）；西北农林科技大学试验示范站(基地)科技成果推广（TGZX2015-32）；陕西省科技统筹创新工程计划（2016KTZDNY02-05）。 |
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| 中文摘要:根据GenBank已登录的PPRV基因组序列，设计并合成2对特异性引物，以PPRV疫苗株基因组为模板，经RT-PCR扩增获得N和M基因并进行序列分析，再将2个基因插入到原核表达载体pET-28a中，经鉴定正确后转化BL21感受态细胞进行IPTG诱导表达，并进行SDS-PAGE和Western blot检测。成功克隆PPRV疫苗株N和M基因，重组菌经IPTG诱导后表达分子质量为57.7 ku N蛋白（核衣壳蛋白）和38.1 ku的M蛋白（基质蛋白），2个蛋白均能与小反刍兽疫阳性血清特异性结合，具有良好的反应原性。 |
| 中文关键词:小反刍兽疫毒 N基因 M基因 克隆 序列分析 原核表达 |
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| Cloning,Sequence Analysis and Prokaryotic Expression of N and M Gene of Peste des Petits Ruminants Virus Vaccine Strain |
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| Abstract:According to the published sequence of N and M gene of PPRV Nigeria 75/1 vaccine strain，two pairs of specific primers were designed and synthesized. N and M gene were amplified by RT- PCR from PPRV. N and M gene sequences were analyzed and cloned into pET-28a vector. The recombinational prokaryotic expression plasmid pET28a-N and pET28a-M were successfully constructed. E.coli BL21 transformed by recombinant plasmid pET28a-N and pET28a-M were induced by IPTG and detected by SDS-PACE and Western blot analysis. N and M gene were successfully cloned and expressed. SDS-PAGE analysis showed that 57.7 ku N and 38.1 ku M fusion protein were expressed by IPTG induction,respectively. Western blot analysis showed that the N and M fusion protein could combine with PPR positive serum and had well reactiongenicity. This experiment provides molecular biology characteristics of PPRV. Further more,the N and M protein can be used to prepare PPRV antibody detection kit. |
| keywords:PPRV N gene M gene Clone Sequence analysis Prokaryotic expression |
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