• 제목/요약/키워드: Recombinant baculovirus

검색결과 158건 처리시간 0.032초

Construction of a Novel Recombinant Bombyx mori Nuclear Polyhedrosis Virus Producing the Fluorescent Polyhedra

  • Kang, Seok-Woo;Yun, Eun-Young;Woo, Soo-Dong;Goo, Tae-Won;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • 제3권1호
    • /
    • pp.75-81
    • /
    • 2001
  • We have constructed a novel recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV) producing the green fluorescent polyhedra. For the production of the fluorescent polyhedra, partial polyhedrin gene containing KRKK as nuclear localization site from the BmNPV polyhedrin gene and the green fluorescent protein (gfp) gene were introduced under the control of p10 promoter of BmNPV. The recombinant BmNPV was stably produced fluorescent polyhedra in the infected Bm5 cells and the morphology of the fluorescent polyhedra was similar to that of wild-type BmNPV. The fluorescent polyhedra had 32 kDa native polyhedrin and 41 kDa fusion protein. From these data, we have further developed a novel BmNPV p10-based transfer vector producing recombinant polyhedra with foreign gene Product. The novel BmNPV P10-based transfer vector is composed of partial polyhedrin gene, factor Xa, and multiple cloning sites.

  • PDF

Substrate Specificity of Human Flavin-containing monooxygenase 1 for Thiocarbamides

  • Jung, Ki-Hwa;D. M. Ziegler;Kim, Young-Mi
    • 한국환경독성학회:학술대회논문집
    • /
    • 한국환경독성학회 2001년도 춘계심포지움 및 학술발표회
    • /
    • pp.124-124
    • /
    • 2001
  • Microsomes isolated from Spodoptera frugiperda (Sf)9 cells infected wi th human FM01 recombinant baculovirus catalyzed the NADPH- and 02-dependent oxidation of methimazole, thiourea, and phenylthiourea. However there was no detectable activity with 1,3-diphenylthiourea or larger thiocarbamides. Microsomes from control Sf9 cells were devoid of methimazole or thiourea S-oxygenase activity. (omitted)

  • PDF

곤충세포주에서 Autographa californica 핵다각체병 바이러스의 다각체 단백질과 초록색 형광 단백질의 융합단백질 발현 및 특성 (Expression and Characterization of Fusion Protein with Autographa californica Nuclear Polyhedrosis Virus Polyhedrin and Green Fluorescent Protein in Insect Cells)

  • 제연호;진병래;노종열;장진희;강석권
    • 한국응용곤충학회지
    • /
    • 제38권2호
    • /
    • pp.139-144
    • /
    • 1999
  • Autographa californica 핵다각체병 바이러스(AcNPV)의 다각체 단백질과 초록색 형광 단백질의 융합단백질의 특성을 분석하였다. 초록색 형광 단백질 유전자는 AcNPV의 완전한 다각체 단백질 유전자의 앞쪽과 뒤쪽에 융합하여 다각체 단백질 유전자의 프로모터 조절하에 도입하였다. 이렇게 작성된 재조합 바이러스를 각각 Ac-GFPPOL 또는 Ac-POLGFP이라고 명명하였다. 이들 재조합 바이러스에 의해 감염된 곤충세포주에서는 56kDa의 융합단백질이 발현되었다. 한편, 흥미롭게도 재조합 바이러스 Ac-POLGFP에 의해 감염된 세포주에서는 초록색 형광이 핵내에서만 다각체 유사 granular particle 형태로 관찰되었다. 반면에 Ac-GFPPOP에 의해 감염된 세포도주에서는 대부분 핵내에 존재하였지만, 세포질과 핵 모두에서 초록색 형광을 관찰할 수 있었다. 그러나 발현된 융합단백질은 분명히 다각체단백질을 포함하고 있음에도 다각체는 형성하지 않았다. 이러한 결과들은 융합단백질에서 다각체단백질의 위치와 관련이 있는 것으로 보여진다.

  • PDF

Overexpression, Purification, and Characterization of the Herpes Simplex Virus-1 DNA Polymerase-UL42 Protein Complex

  • Song, Byeong-Doo;Lehman, I. Robert
    • BMB Reports
    • /
    • 제31권6호
    • /
    • pp.585-589
    • /
    • 1998
  • The herpes simplex virus type-1 (HSV-1)-encoded DNA polymerase consists of two subunits, the products of the UL30 and UL42 genes. UL30 and UL42 were coexpressed in Sf9 cells infected with recombinant baculoviruses carrying the two genes. The UL30 and UL42 gene products remained tightly associated throughout the purification, which led to a near homogeneous heterodimer composed of the DNA polymerase and UL42 protein. The DNA polymerase-UL42 protein heterodimer, purified from the recombinant baculovirus-infected Sf9 cells, showed the same high degree of processivity of deoxynucleotide polymerization as the enzyme purified from the HSV-1 infected primate cells. Like the latter, it contained a 3'-5' exonuclease activity that specifically hydrolyzes an incorrectly matched nucleotide at the 3' terminus of a primer, thereby contributing to the fidelity of DNA replication.

  • PDF