대장균에서 돼지 생식기ㆍ 호흡기 증후군 바이러스 ORF 7 유전자의 발현

Expression of the PRRS Virus ORF 7 Gene in E. coli

As a preceding study for the development of a safe and new recombinant vaccine or a diagnostic kit against Porcine reproductive and respiratory syndrome (PRRS), we plan to produce an open reading frame 7 (ORF 7) protein of PRRS virus. To do this, we construct a pET-ORF 7 recombinant expression vector by cloning the ORF 7 gene into the pET expression vector. Then the expression vector pET-ORF 7 was transformed into Escherichia coli BL21/DE3 and expression was induced by Isopropyl β-D-1-thiogalactopyranoside (IPTG). The relative amount of the ORF 7 protein was analyzed by SDS-PAGE. The recombinant plasmid pET-ORF 7 was identified to be expressed successfully in E. coli. The molecular weight of the expression protein was approximate 14 kDa, The ORF 7 protein was detected at 30 min after IPTG induction and increased to 4 hours as time-dependent manner. We will utilize it for the preparation of monoclonal antibody necessary to the development of a diagnostic kit against PRRS virus.