Baculovirus Expression Vectors: the Requirements for High Level Expression of Proteins, Including Glycoproteins

Matsuura, Yoshiharu; Possee, Robert D.; Overton, Hilary A.; Bishop, David H. L.

doi:10.1099/0022-1317-68-5-1233

Cited by 669 publications

(388 citation statements)

References 23 publications

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“…Complementary DNA of human H-Ras[G12V] was subcloned into the BamHI sites of a baculovirus transfer vector, pAcYM1 (Matsuura et al, 1987) (a kind gift from Yoshiharu Matsuura, National Institute of Health, Japan) to obtain pAcYM1-Ras[G12V]. Plasmid pLNC-Raf-1 : FH6 and bacterial expression system for recombinant histidinetagged Xenopus MEK (His-MEK) and glutathione-S-transferase ± fused kinase-de®cient Xenopus MAPK (GST-kdMAPK) (Kosako et al, 1993), were kindly given by Martin McMahon (DNAX Research Institute, Palo Alto, CA) and Eisuke Nishida (Kyoto University, Japan), respectively.…”

Section: Methodsmentioning

confidence: 99%

Isolation of a new protein factor required for activation of Raf-1 by Ha-Ras: partial purification from rat brain cytosols

1998

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Ras-mediated signaling pathways play a critical role in cellular proliferation and di erentiation. Although it has been demonstrated that Ras interacts with Raf-1 to stimulate the serine/threonine kinase activity of Raf-1, the precise mechanism by which Ras activates Raf-1 remains obscure. To address this question, we developed a cell-free system in which the activated form of H-Ras can induce Raf-1 activation. Using this system, we found the presence of a new protein factor, in cytosolic fractions of both human embryonic kidney 293 cells and rat brain tissues, that is required for Ras-dependent activation of Raf-1. The factor was puri®ed from rat brain cytosols through successive column chromatographies on DEAE Sephacel, SP Sepharose and Sephacryl S-300. The approximate molecular weight of the activator was estimated as 400 000 by gel ®ltration. Its activity was sensitive to heat and trypsin treatments. The puri®ed activator did not contain Src, 14-3-3, protein kinase C, JAK2 or Ksr-1, as judged by immunoblotting. Further characterization of the activator is underway.

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Section: Methodsmentioning

confidence: 99%

Isolation of a new protein factor required for activation of Raf-1 by Ha-Ras: partial purification from rat brain cytosols

1998

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“…Spodoptera frugiperda clonal isolated 9 (SIX)) cells, vector plasmid pAcYM1 [9], and wild-type Autgrapha californica nuclear polyhedrosis virus (AcNPV) were supplied by Dr. A. Takase (National Institute of Neuroscience, N.C.N.P., Tokyo, Japan). Egg phosphatidylcholine and dl-ctotocopherol were from Sigma (Deisenhofen, Germany).…”

Section: Methodsmentioning

confidence: 99%

Only sphingolipid activator protein B (SAP‐B or saposin B) stimulates the degradation of globotriaosylceramide by recombinant human lysosomal α‐galactosidase in a detergent‐free liposomal system

et al. 1996

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The degradation of globotriaosylceramide (GbOse3Cer) by insect-cell derived recombinant human ct-galactosidase (EC 3.2.1.22) was carried out in a detergent-free iiposomal system in order to mimic intralysosomal conditions. GbOse3Cer incorporated into unilamellar liposomes was used as the substrate, and naturally occurring sphingolipid activator proteins, rather than detergents, were used to stimulate the enzyme reaction. The degradation of GbOse3Cer was dependent on the presence of both ct-galactosidase and sphingolipid activator protein B (SAP-B or saposin B). It proceeded optimally at pH 4.6, and was enhanced by increasing amounts of both ctgalactosidase (0.24-24 mU/50 lal assay) and SAP-B (0-5 pg/50 p] assay). The enzyme reaction was not affected by SAP-A, SAP-C, or SAP-D. Therefore, our results indicate that only SAP-B is essential for the degradation of GbOse3Cer by ctgalactosidase.

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“…With the recent development of baculovirus expression systems, the production of large amounts of viral antigen produced in Spodoptera frugiperda (Sf9) and other insect cells in biocontainment-free environments has become possible. In this system, Autographa californica nuclear polyhedrosis virus (baculovirus) can be genetically altered to express unusually high levels of foreign gene products when the gene is inserted into the baculovirus polyhedron gene (69,77). Utilization of this technique to produce large amounts of flavivinas antigen to be used in diagnostic tests may ultimately reduce the need for containment facilities in diagnostic laboratories.…”

Section: Characteristics Of Flavivirus Replication In Cultured Cellsmentioning

confidence: 99%

Insect-transmitted vertebrate viruses: Flaviviridae

Ludwig

Iacono-Connors

1993

In Vitro Cell Dev Biol - Animal

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SUMMARYThe Flaviviridae include almost 70 viruses, nearly half of which have been associated with human disease. These viruses are among the most important arthropod-borne viruses worldwide and include dengue, yellow fever, and Japanese encephalitis viruses. Morbidity and mortality caused by these viruses vary, but collectively they account for millions of encephalitis, hemorrhagic fever, arthralgia, rash, and fever cases per year. Most of the members of this family are transmitted between vertebrate hosts by arthropod vectors, most commonly mosquitoes or ticks. Transmission cycles can be simple or complex depending on the hosts, vectors, the virus, and the environmental factors affecting both hosts and viruses. Replication of virus in invertebrate hosts does not seem to result in any significant pathology, which suggests a close evolutionary relationship between virus and vector. Another example of this relationship is the ability of these viruses to grow in invertebrate ceil culture, where replication usually results in a steady state, persistent infection, often without cytopathic effect. Yields of virus from insect cell culture vary but are generally similar to yields in vertebrate cells. Replication kinetics are comparable between insect and vertebrate ceil lines, despite differences in incubation temperature. Both vertebrate and insect cell culture systems continue to play a significant role in flavivirus isolation and the diagnosis of disease caused by these agents. Additionally, these culture systems permit the study of flavivirus attachment, penetration, replication, and release from cells and have been instrumental in the production and characterization of live-attenuated vaccines. Both vertebrate and insect cell culture systems will continue to play a significant role in basic and applied flavivirus research in the future.

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Baculovirus Expression Vectors: the Requirements for High Level Expression of Proteins, Including Glycoproteins

Cited by 669 publications

References 23 publications

Isolation of a new protein factor required for activation of Raf-1 by Ha-Ras: partial purification from rat brain cytosols

Isolation of a new protein factor required for activation of Raf-1 by Ha-Ras: partial purification from rat brain cytosols

Only sphingolipid activator protein B (SAP‐B or saposin B) stimulates the degradation of globotriaosylceramide by recombinant human lysosomal α‐galactosidase in a detergent‐free liposomal system

Insect-transmitted vertebrate viruses: Flaviviridae

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