Tyrosine kinase B (Trk B) 재조합 단백질과 특이 항체 생산

Abstract

Tyrosine kinase B (Trk B) is the intracellular receptor for the brain-derived neurotrophic factor (BDNF). In order to study the relationship between the neurotrophin BDNF and its receptor Trk B, the productions of recombinant Trk B proteins were established and the following results were obtained. A partial Trk B gene of rat brain was cloned into E. coli expression vector under the control of T7 promoter. When these partial cDNAs were expressed as recombinant partial Trk B proteins by addition of IPTG, they were accounted for about 20% of total E, coli proteins, displaying a molecular size of 38KDa. Partial Trk B were purified and refolded by metal affinity chromatography and dialysis showing 38KDa as monomer and 76KDa as dimer, respectively. After three times immunizations of rabbit through six weeks, the polyclonal antisera were separated from the blood, and their specificities and titers were investigated by ELISA and immunohistology, respectively.