• Title/Summary/Keyword: Baculovirus

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High-efficient Expression of Porcine IL-2 with Recombinant Baculovirus Infected Silkworm, Bombyx mori

  • Inumaru, Shigeki;KokuHo, Takehiro;Yada, Takashi;Kiuchi, Makoto;Miyazawa, Mitsusuhiro
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.146-149
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    • 2000
  • Biologically active porcine Interleukin-2(poIL-2) was produced from in vitro and in viva baculovirus expression system, namely the Autographa californica nuclear polyhedrosis virus (ACNPV)-cell culture system and the Hybrid nucler polthedrosis virus (HyNPV)-sillkworm larva system. The concentration of the recombinant poIL-2(rpoIL-2) in the larvae hemolymph was 1 to 3 mg/mL, which was about 7 to 20 times those of the cell culture systems. The level of this expression efficiency is equal to that with transgenic livestock, secretion products in milk.

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Diagnosis of Viral Infection of Pensaeide (보리새우류의 바이러스 감염증의 진단)

  • 허문수;정초록;장경립
    • Journal of Life Science
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    • v.9 no.4
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    • pp.453-459
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    • 1999
  • Baculovirus(WSBV) was isolated from infected Penaeide was collected from shrimp farm at southern sea of Korea from 1993 to 1995. The Infectious virus was purified and used for diagnosis of infected shrimp. Anti-viral serum were used for immunological detection as enzyme linked immunoabsorbent assay(ELISA) and indirect fluorescent antibody technique(IFAT). In IFAT, stomach, lymphoid organ and antenae gland of infected shrimp showed fluorescent reaction. In ELISA, tissues of spontaneously infected shrimp appeared higher O.D. values than in artificial infected shrimp. Primer set was constructed from sequence of 420bp of cloned Baculovirus(WSBV) genome. Specific band for infected shrimp was detected in Polymerase chain reaction(PCR)

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Estimation of Human Flavin-containing Monooxygenases Activity(FMO1) in the Baculovirus Expression Vector System by using S-oxidation of Methimazole

  • Kim, Young-Mi
    • Journal of Food Hygiene and Safety
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    • v.14 no.4
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    • pp.415-421
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    • 1999
  • The flavin-containing monooxygenases (FMOs) (EC 1.14. 13.8) are NADPH-dependent flavoenzymes that catalyze oxidation of soft nucleophilic heteroatom centers in a range of structurally diverse compounds including foods, drugs, pesticides, and other xenobiotics. In humans, FMOl appears to be the predominant form expressed in human fetal liver. cDNA-expressed human FMO and human liver microsomal FMO have been observed to N- and S-oxy-genate nucleophilic nitrogen- and sulfur-containing drugs and chemicals, respectively. In the present study, FMOl can be expressed in the baculovirus expression vector system at level of 2.68 nmol FMOl/mg of membrane protein. This isoform was examined for its capacity to metabolize methimazole to its S-oxide using thiocholine assay. Kinetic studies of its S-oxide by recombinant human FMO1 result in Km of 7.66 $\mu$M and Vmax of 17.79 nmol/min/mg protein.

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Comparison of enzymatic activities between the recombinant CHT1 proteins from the hard tick Haemaphysalis longicornis expressed in E. coli and baculovirus-mediated Sf 9 cells (E. coli와 baculovirus-mediated Sf 9 세포에서 발현된 진드기 H. longicornis의 CHT1 단백의 효소활성 비교)

  • You, Myung-Jo;Fujisaki, Kozo
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.139-144
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    • 2003
  • A chitinase cDNA named CHT1 was cloned from the hard tick, Haemaphysalis longicornis, and the enzymatic properties of its recombinant proteins were characterized. The CHT1 cDNA encodes 930 amino-acid (aa) residues including a 22 aa putative signal peptide, with the calculated molecular mass of the putative mature protein 104 kDa. The E coli-expressed rCHT1 exhibited weak chitinolytic activity against $4MU-(GlcNAc)_3$. The rCHT1 protein with higher activity was obtained using recombinant Autographa californica multiple nuclear polyhedrosis virus (AcMNPV), which expresses rCHT1 under polyhedrin promoter. These findings suggest that the rCHT1 expressed in baculovirus-mediated Sf 9 cells has a high activity than E coli-expressed rCHT1.

White Spot Baculovirus Infection of Shrimp Spawner, Inhabitant Organisms and Survival in Seawater (White Spot Baculovirus의 모하와 중간숙주 감염 및 해수생존)

  • Heo, Moon-Soo;Sohn, Sang-Gyu;Kim, Young-Jin
    • Journal of fish pathology
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    • v.13 no.1
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    • pp.15-20
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    • 2000
  • Infection rates with white spot baculovirus(WSBV) in wild-caught shrimp spawners appeared to be 52% in Penaeus chinensis and 20% in Penaeus japonicus when diagnosed by polymerase chain reaction (PCR). Infection rates of WSBV from inhabitant organisms in shrimp farm showed 37% in Upogebia major and 29% in Calliamassa japonica. The viruses maintained their survivorship in seawater at $4^{\circ}C$, but lost it at $25^{\circ}C$.

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Insect cell-baculovirus system을 이용한 $TGF-{\beta}_1$의 최적 생산전략

  • Lee, Chang-Jin;Chae, Jong-Seok;Cha, Sang-Hun;Jeon, Gye-Taek;Jeong, Yeon-Ho
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.396-397
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    • 2000
  • $TGF-{\beta}_1$을 insect cell-baculovirus system을 이용하여 생산하였으며, 이 경우 낮은 세포밀도와 암모니아 같은 노폐물에 의한 생산성 저해 문제를 해결하기 위해 유가식 배양과 흡착제에 의한 암모니아 제거를 동시에 수행함으로써 $TGF-{\beta}_1$의 생산성을 향상시킬 수 있었다.

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Substrate Specificity of the Human Flavin-containing Monooxygenase for Organic Selenium Compounds (사람 Flavin-containing Monooxygenase의 셀레니움화합물에 대한 기질 특이성에 관한 연구)

  • Kim, Young-Mi
    • Environmental Analysis Health and Toxicology
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    • v.15 no.4
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    • pp.139-145
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    • 2000
  • The Flavin-containing monooxygenase (FMOs) (EC1.14.13.8) are NADPH-dependent flavoenzymes that catalyze oxidation of soft nucleophilic heteroatom centers in a range of structurally diverse compounds, including foods, drugs, pesticides, and other xenobiotics. In humans, FMO3 is quantitatively a major human liver monooxygenase. In the present study, the baculovirus expression vector system was used to overexpress human FMO3 in insect cells for catalytic studies. Six commercially available organic selenium compounds were examined for substrate activity with microsomes isolated from Spodoptera frugiperda (Sf)9 cells infected with human FMO3 recombinant baculovirus. While none of the aromatic heterocyclic selenides tested showed detectable activity, all dialkyl- and alkylaryl-selenides free from ionic groups catalyzed the NADPH- and O$_2$-dependent oxidation. Kinetic constants demonstrate that (based on Km) dialkyl-and alkylaryl- selenides are better substrates for human FMO3 than analogous nitrogen or sulfur compounds .

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Effect of Substrate Size on Activities of Thiocarbamides with the Human Flavin-containing Monooxygenase 3 (사람 Flavin-containing Monooxygenase 3의 Thiocarbamide 화합물의 기질 크기에 따른 효소활성에 관한 연구)

  • 김영미
    • Environmental Analysis Health and Toxicology
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    • v.16 no.2
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    • pp.97-102
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    • 2001
  • The flavin-containing monooxygenases(FMOs) (EC1.14.13.8) are NADPH0dependent flavoenzymes that catalyze oxidation of soft nucleophilic heteroatom centers in a range of structurally diverse compounds, including foods, drugs, pesticides, and other xenobiotics. In humans, FMO3 is quantitatively a major human liver monooxygenase. In the present study, the baculovirus expression vector system was used to overexpress human FMO3 in sect cells for stalytic studies. Microsomes isolated from Spodoptera frugiperda(Sf)9 cells infected with human FMO3 recombinant baculovirus catalyzed the NADPH-and O$_2$-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. 1, 3-diphenylthiourea is apparently completely excluded from the catalytic site, these amines drugs are probably approaching the upper size limits of xenobiotics accepted by human FMO3. The substrate specificity of this iosform in humans appears considerably more restriceted than that of pig, guinea pig, rat or rabbit FMO3.

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