Expression and Characterization of Fusion Protein with Autographa californica Nuclear Polyhedrosis Virus Polyhedrin and Green Fluorescent Protein in Insect Cells

곤충세포주에서 Autographa californica 핵다각체병 바이러스의 다각체 단백질과 초록색 형광 단백질의 융합단백질 발현 및 특성

  • 제연호 (서울대학교 농업생명과학대학 응용생물화학부 및 농업생물신소재 연구센터) ;
  • 진병래 (동아대학교 생명자원과학대학) ;
  • 노종열 (서울대학교 농업생명과학대학 응용생물화학부 및 농업생물신소재 연구센터) ;
  • 장진희 (서울대학교 농업생명과학대학 응용생물화학부 및 농업생물신소재 연구센터) ;
  • 강석권 (서울대학교 농업생명과학대학 응용생물화학부 및 농업생물신소재 연구센터)
  • Published : 1999.08.01

Abstract

We have now constructed a novel recombinant baculovirus producing fusion protein with Autogrqha c.uliforrzica nuclear polyhedrosis virus (AcNPV) polyhedrin and green fluorescent protein (GFP). The fusion protein expressed by the recombinant baculovirus in insect cells was characterized. The GFP gene was introduced under the control of polyhedrin gene promoter of AcNPV, by fusion in the front or back of intact polyhedrin gene. The recombinant baculoviruses were named as Ac-GFPPOL or Ac-POLGFP. respectively. As expected, the 56 kDa fusion protein was expressed in the recombinant virus-infected cells. Interestingly. however, the fluorescence of GFP in the cells infected with Ac- POLGFP was only detected within the nuclei. and that was observed as polyhedra-like granular particles. In the microscopy of cells infected with Ac-GFPPOL, furthermore, GFP was detected in both cytoplasm and nuclei although most of GFP were present within the nuclei. However, fusion protein produced by recombinant virus did not form polyhedra although the fusion protein was fused with polyhedrin and GFP. It is suggested that difference of GFP location in the infected cells appear to be involved in the region of polyhedrin in the fusion protein, and the polyhedrin in the fusion protein might be responsible for the polyhedra-like granular particles present within nuclei.