Prokaryotic soluble expression method of PRRSV N protein, and indirect ELISA antibody detection method

• Main Authors: LYU JIANLIANG, DAI JUNFEI, ZHANG JIE, OU YUNWEN, ZHANG YONGGUANG, WANG JUN, MA BING, LIU YONGSHENG, SHAO JUNJUN, ZHOU PENG, DING YAOZHONG, SUN YUEFENG
• Format: Patent
• Language: Chinese; English
• Published: 07.09.2018
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PHYSICS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; TESTING; VINEGAR; WINE

The invention discloses a prokaryotic soluble expression method of a PRRSV N protein, and an indirect ELISA antibody detection method. The prokaryotic soluble expression method of the PRRSV N proteincomprises the following steps: (1) artificially optimizing and synthesizing a PRRSV N protein encoding gene according to the codon bias characteristics of Escherichia coli; (2) constructing a prokaryotic expression vector of an N protein by using MBP as a solubilizing tag; and (3) expressing and purifying an MBP fusion N protein, and performing specificity test by the indirect ELISA antibody detection method. The high-efficiency soluble expression of the PRRSV N protein in Escherichia coli is realized by using PMAL-C4X with the MBP tag as an expression vector, induced culturing is carried outat a temperature close to room temperature without a heating or cooling device, and the target protein is eluted with a 10 mmoL/L maltose solution, so good elution effect, low dosage and cost saving are achieved. The indirect