The open reading frame (ORF) of hAPOA1 was inserted into the prokaryotic expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX- 4T-I-hAPOA1, which was then transformed into Escherichia coil strain BL...The open reading frame (ORF) of hAPOA1 was inserted into the prokaryotic expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX- 4T-I-hAPOA1, which was then transformed into Escherichia coil strain BL21. The expression of target fusion protein was induced with isopropyl β-D-l-thiogalacto- pyranoside (IPTG). The purified fusion protein in inclusion bodies was used to immunize New Zealand white rabbits to prepare hAPOA1 antiserum and the antibody titer was detected with indirect enzyme-linked immunosorbent assay (ID-EL1SA). ID-ELISA and Western Blot proved that rabbit polyclonal antibody with a high titer of 1 : 40 000 was produced, which may bring considerable economic benefits.展开更多
基金Supported by the Agricultural Science Independent Innovation Fund Jiangsu Province[CX(16)1326]
文摘The open reading frame (ORF) of hAPOA1 was inserted into the prokaryotic expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX- 4T-I-hAPOA1, which was then transformed into Escherichia coil strain BL21. The expression of target fusion protein was induced with isopropyl β-D-l-thiogalacto- pyranoside (IPTG). The purified fusion protein in inclusion bodies was used to immunize New Zealand white rabbits to prepare hAPOA1 antiserum and the antibody titer was detected with indirect enzyme-linked immunosorbent assay (ID-EL1SA). ID-ELISA and Western Blot proved that rabbit polyclonal antibody with a high titer of 1 : 40 000 was produced, which may bring considerable economic benefits.