Citation: . 猪繁殖与呼吸综合征病毒N基因的克隆及高效表达* .VIROLOGICA SINICA, 2003, 18(3) : 279-282.

猪繁殖与呼吸综合征病毒N基因的克隆及高效表达*

  • Available online: 18 June 2003
  • The gene of nucleocapsid protein of Porcine reproductive and respiratory syndrome virus (PRRSV) VR2332 strain was amplified by RT-PCR. The PCR product was purified and digested with BamH Ⅰ and Xho Ⅰ, then directly cloned into the prokaryotic vector pET32a. Consequently the recombinant plasmid was constructed, designated pETN. PETN was transformed into the host cell BL21 (DE3) and the expression procedure was optimized including cultivated temperature, optional induction concentration and time of IPTG. The result indicated that the nucleocapsid protein can be expressed efficiently with 0.8mmol/L IPTG and 4-hour induction. The gene of nucleocapsid protein of Porcine reproductive and respiratory syndrome virus (PRRSV) VR2332 strain was amplified by RT-PCR. The PCR product was purified and digested with BamH Ⅰ and Xho Ⅰ, then directly cloned into the prokaryotic vector pET32a. Consequently the recombinant plasmid was constructed, designated pETN. PETN was transformed into the host cell BL21 (DE3) and the expression procedure was optimized including cultivated temperature, optional induction concentration and time of IPTG. The result indicated that the nucleocapsid protein can be expressed efficiently with 0.8mmol/L IPTG and 4-hour induction.

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    猪繁殖与呼吸综合征病毒N基因的克隆及高效表达*

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    Abstract: The gene of nucleocapsid protein of Porcine reproductive and respiratory syndrome virus (PRRSV) VR2332 strain was amplified by RT-PCR. The PCR product was purified and digested with BamH Ⅰ and Xho Ⅰ, then directly cloned into the prokaryotic vector pET32a. Consequently the recombinant plasmid was constructed, designated pETN. PETN was transformed into the host cell BL21 (DE3) and the expression procedure was optimized including cultivated temperature, optional induction concentration and time of IPTG. The result indicated that the nucleocapsid protein can be expressed efficiently with 0.8mmol/L IPTG and 4-hour induction. The gene of nucleocapsid protein of Porcine reproductive and respiratory syndrome virus (PRRSV) VR2332 strain was amplified by RT-PCR. The PCR product was purified and digested with BamH Ⅰ and Xho Ⅰ, then directly cloned into the prokaryotic vector pET32a. Consequently the recombinant plasmid was constructed, designated pETN. PETN was transformed into the host cell BL21 (DE3) and the expression procedure was optimized including cultivated temperature, optional induction concentration and time of IPTG. The result indicated that the nucleocapsid protein can be expressed efficiently with 0.8mmol/L IPTG and 4-hour induction.

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