Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli

Luckow, Verne A.; Lee, S C; Barry, Gerard F.; Olins, Peter O.

doi:10.1128/jvi.67.8.4566-4579.1993

Cited by 805 publications

(329 citation statements)

References 45 publications

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“…However, the final virus stock unavoidably contains a mixture of parental and recombinant viruses, so a plaque-assay is required to purify the recombinant baculovirus [10]. An alternative approach has been developed [11] to circumvent this problem by generating a recombinant baculovirus using site-specific transposition with Tn7 to insert foreign genes into bacmid (baculovirus plasmid) propagated in E. coli. Based on the principle of site-specific transposition, a rapid and efficient BES (Bac-to-Bac Ò , Life Technologies) that can be used as an alternative way to generate recombinant baculoviruses was developed and is now widely used as another commercial BES.…”

Section: Bes As a Platform For Protein Expressionmentioning

confidence: 99%

Use of baculovirus expression system for generation of virus-like particles: Successes and challenges

Liu

et al. 2013

Protein Expression and Purification

106

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The baculovirus expression system (BES) has been one of the versatile platforms for the production of recombinant proteins requiring multiple post-translational modifications, such as folding, oligomerization, phosphorylation, glycosylation, acylation, disulfide bond formation and proteolytic cleavage. Advances in recombinant DNA technology have facilitated application of the BES, and made it possible to express multiple proteins simultaneously in a single infection and to produce multimeric proteins sharing functional similarity with their natural analogs. Therefore, the BES has been used for the production of recombinant proteins and the construction of virus-like particles (VLPs), as well as for the development of subunit vaccines, including VLP-based vaccines. The VLP, which consists of one or more structural proteins but no viral genome, resembles the authentic virion but cannot replicate in cells. The high-quality recombinant protein expression and post-translational modifications obtained with the BES, along with its capacity to produce multiple proteins, imply that it is ideally suited to VLP production. In this article, we critically review the pros and cons of using the BES as a platform to produce both enveloped and non-enveloped VLPs.

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Section: Bes As a Platform For Protein Expressionmentioning

confidence: 99%

Use of baculovirus expression system for generation of virus-like particles: Successes and challenges

Liu

et al. 2013

Protein Expression and Purification

106

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“…They are based on the E. coli fertility factor (F-factor) replicon which is maintained as a circular supercoiled extra chromosomal single copy plasmid in the bacterial host (Monaco and Larin, 1994;O'Connor et al, 1989;Shizuya et al, 1992). Their high capacity suited their primary use for the construction of infectious clones of large DNA viruses such as baculoviruses (Luckow et al, 1993) or herpesvirus (Saeki et al, 1998).…”

Section: Figmentioning

confidence: 99%

Flavivirus reverse genetic systems, construction techniques and applications: A historical perspective

et al. 2015

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The study of flaviviruses, which cause some of the most important emerging tropical and sub-tropical human arbovirus diseases, has greatly benefited from the use of reverse genetic systems since its first development for yellow fever virus in 1989. Reverse genetics technology has completely revolutionized the study of these viruses, making it possible to manipulate their genomes and evaluate the direct effects of these changes on their biology and pathogenesis. The most commonly used reverse genetics system is the infectious clone technology. Whilst flavivirus infectious clones provide a powerful tool, their construction as full-length cDNA molecules in bacterial vectors can be problematic, laborious and time consuming, because they are often unstable, contain unwanted induced substitutions and may be toxic for bacteria due to viral protein expression. The incredible technological advances that have been made during the past 30years, such as the use of PCR or new sequencing methods, have allowed the development of new approaches to improve preexisting systems or elaborate new strategies that overcome these problems. This review summarizes the evolution and major technical breakthroughs in the development of flavivirus reverse genetics technologies and their application to the further understanding and control of these viruses and their diseases.

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“…At about the same time, another efficient and rapid method for generation of recombinant baculoviruses was developed (Luckow et al, 1993) that employed transposition of a foreign gene expression cassette from a donor plasmid into a bacterial artificial chromosome (BAC) which contains the entire AcMNPV genome (bacmid). In this system (Bac-to-Bac TM ) recombinant baculovirus genomes are generated in Escherichia coli and then used to transfect insect cells to obtain recombinant baculovirus particles.…”

Section: B Baculovirus Vectorsmentioning

confidence: 99%

Vaccines for Viral and Parasitic Diseases Produced with Baculovirus Vectors

Oers

2006

Advances in Virus Research

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The baculovirus-insect cell expression system is an approved system for the production of viral antigens with vaccine potential for humans and animals and has been used for production of subunit vaccines against parasitic diseases as well. Many candidate subunit vaccines have been expressed in this system and immunization commonly led to protective immunity against pathogen challenge. The first vaccines produced in insect cells for animal use are now on the market. This chapter deals with the tailoring of the baculovirus-insect cell expression system for vaccine production in terms of expression levels, integrity and immunogenicity of recombinant proteins, and baculovirus genome stability. Various expression strategies are discussed including chimeric, virus-like particles, baculovirus display of foreign antigens on budded virions or in occlusion bodies, and specialized baculovirus vectors with mammalian promoters that express the antigen in the immunized individual. A historical overview shows the wide variety of viral (glyco)proteins that have successfully been expressed in this system for vaccine purposes. The potential of this expression system for antiparasite vaccines is illustrated. The combination of subunit vaccines and marker tests, both based on antigens expressed in insect cells, provides a powerful tool to combat disease and to monitor infectious agents.

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Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli

Cited by 805 publications

References 45 publications

Use of baculovirus expression system for generation of virus-like particles: Successes and challenges

Use of baculovirus expression system for generation of virus-like particles: Successes and challenges

Flavivirus reverse genetic systems, construction techniques and applications: A historical perspective

Vaccines for Viral and Parasitic Diseases Produced with Baculovirus Vectors

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