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
根据鸡缩胆囊素(choleystok in in,cck)-33基因的碱基序列及大肠杆菌高频密码子,设计了大肠杆菌表达系统偏爱的cck基因序列.将cck-33基因克隆至pRSET A质粒上,利用载体上的同尾酶构建cck基因四串联体并在大肠杆菌E.coliBL 21中成功表达...根据鸡缩胆囊素(choleystok in in,cck)-33基因的碱基序列及大肠杆菌高频密码子,设计了大肠杆菌表达系统偏爱的cck基因序列.将cck-33基因克隆至pRSET A质粒上,利用载体上的同尾酶构建cck基因四串联体并在大肠杆菌E.coliBL 21中成功表达.将所表达的融合蛋白进行纯化后,以纯化的蛋白为免疫原制备油佐剂疫苗主动免疫胡须肉鸡.结果表明,CCK蛋白主动免疫肉鸡后,抗血清水平显著升高,P/N值远远大于2.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.
文摘根据鸡缩胆囊素(choleystok in in,cck)-33基因的碱基序列及大肠杆菌高频密码子,设计了大肠杆菌表达系统偏爱的cck基因序列.将cck-33基因克隆至pRSET A质粒上,利用载体上的同尾酶构建cck基因四串联体并在大肠杆菌E.coliBL 21中成功表达.将所表达的融合蛋白进行纯化后,以纯化的蛋白为免疫原制备油佐剂疫苗主动免疫胡须肉鸡.结果表明,CCK蛋白主动免疫肉鸡后,抗血清水平显著升高,P/N值远远大于2.
