Baculovirus를 이용한 Aujeszky's Disease Virus gⅢ 단백질 발현 = Glycoprotein gⅢ of Aujesky's Disease Virus Expressed in Insect Cells by a Baculovirus

다국어 초록 (Multilingual Abstract)

The g Ⅲ gene located in U_L region of Yangsan strain, a field isolate of Aujeszky's disease virus (ADV) in Korea, was cloned into pTZ18R and sequenced. The gⅢ gene consisting of 1,437 nucleotides showed 98% sequence homology with that of Becker strain, a reference strain of ADV.
The gene encoding gⅢ of Yangsan strain was placed under the control of Autographa californica nucleopolyhedrosis virus (AcNPV) polyhedrin promoter, and expressed by the derived recombinant baculovirus using Spodoptera frugiperda 9 (Sf9) cells. The expressed gⅢ was a protein with molecular weight of 72kd determined by immunoprecipitation and Western blotting assay using anti-ADV polyclonal antibodies and anti-gⅢ monoclonal antibody.
The partially purified gⅢ protein was utilized as antigen in the radial immunodiffusion enzyme assay (RIDEA) to detect to specific antibody against ADV in pig sera. The results indicated that the sensitivity of RIDEA with the recombinant gⅢ protein antigen (98%) was as high as that with the conventional glycoprotein antigen extracted from the ADV infected cells. In addition, the false positive and false nagative reactions in gⅢ RIDEA were significantly reduced than the conventional glycoprotein RIDEA as judged from the results of standard serum neutralization test.