微生物学技术

Wave Bioreactors are relatively new to the field of protein production from insect cells infected with recombinant baculoviruses. Various sizes are available to support expression needs from small to large scale. They offer many advantages over stirred tank and airlift reactors, inc ...

With an increasing need for functional analysis of proteins, there is a growing demand for fast and cost-effective production of biologically active eukaryotic proteins. The baculovirus expression vector system is widely used, and in the vast majority of cases cultured insect cells have ...

The ability to make a large variety of virus-like particles (VLPs) has been successfully achieved in the baculovirus expression vector system (BEVS)/insect cell system. The production and scale-up of these particles, which are mostly sought as candidate vaccines, are currently being ad ...

Techniques and experimental applications are described for exogenous protein expression in Drosophila cell lines. Ways in which the Drosophila cell lines and the baculovirus expression vector system differ in their applications are emphasized.

The baculovirus expression vector system is widely used to produce large quantities of recombinant proteins. However, yields of extracellular and membrane-bound proteins obtained with this system often are very low, possibly because of the adverse effects of baculovirus infecti ...

Baculoviruses are widely used both as protein expression vectors and as insect pest control agents. This section provides an overview of the baculovirus lifecycle and use of baculoviruses as insecticidal agents. This chapter includes discussion of the pros and cons for use of baculoviru ...

The lepidopteran insect cells used with the baculovirus expression vector system (BEVS) are capable of synthesizing and accurately processing foreign proteins. However, proteins expressed in baculovirus-infected cells often fail to be completely processed, or are not proces ...

A considerable amount of work has been done during the last 20 yr to genetically enhance the efficacy of baculovirus insecticides. Following construction of a genetically altered baculovirus, laboratory bioassays are used to quantify various parameters of insecticidal activity s ...

Probing the baculovirus infection process is essential in optimizing the recombinant protein production. Typically, researchers monitor the infection process in the stirred tank reactor, which, however, contains a population of cells infected at different times after virus in ...

Through green fluorescent protein (GFP) strategy, facile and fast monitoring and visualization of baculovirus infection in insect cells is possible in vivo. This chapter describes two novel techniques for simple determination of virus titer in the baculovirus expression system u ...

This chapter describes a technique for synthesizing and transfecting double stranded RNA (dsRNA) for RNA interference in Sf-21 cell culture. Transfection with dsRNA only requires 1 h and the cells are usually recovered within 12 h. Suggestions for designing dsRNA are included in the method ...

Apoptosis is a physiological program of cell suicide conserved in invertebrates and vertebrates. Apoptosis is crucial to the normal development of organisms and in tissue homeostasis, by promoting elimination of unwanted cells including damaged- or virusinfected cells. Because ...

Genetically modified baculoviruses can efficiently deliver and express genes in mammalian cells. The major prerequisite for the expression of a gene transferred by baculovirus is its control by a promoter that is active in mammalian cells. This chapter describes methods for producing ...

A protocol for the development of cancer vaccines is presented. The protocol is based upon the long-term in vitro treatment of cancer cells with interferon (IFN)-α to create cancer vaccine cells. This protocol has been used to develop cancer vaccines in mice against B16 melanoma, RM-1 prostate ca ...

Recent studies have revealed that type I interferons (IFNs) are powerful inducers of the differentiation and activation of dendritic cells (DCs). These findings emphasize the importance of these cytokines in linking innate and adaptive immunity, suggesting that effects of type I IFN on D ...

Plasmacytoid dendritic cells (PDCs) are the natural interferon (IFN-α)-producing cells in human peripheral blood that produce vast quantities of IFN-α in response to viral infection and other stimuli. PDCs are a rare cell type, making up less than 0.5% of peripheral blood mononuclear cells. To ...

A defining hallmark of the type 1 interferons (IFNs) is their ability to interfere with virus replication. As such, viruses have evolved diverse mechanisms to subvert the antiviral effects of interferons. Herpes simplex virus type 1 (HSV-1) is a large deoxyribonucleic acid virus best known for ...

Delivery of type I interferon (IFN) subtypes by intramuscular inoculation of mice with a recombinant mammalian expression vector encoding IFN stimulates the immune response. Such immunomodulation drives towards a Th1-like response. The degree of stimulation of the immune respon ...

Gene transfer is a widely used experimental approach to determine the value of specific genes under a variety of conditions. This chapter focuses on the expression of two human antiviral genes, interferon (IFN)-α2 and IFN-β, driven by a cytomegalovirus immediate-early promoter in a human M�l ...

The cytokine family of interferons (IFNs) has multiple functions, including antiviral, antitumor, and immunomodulatory effects and regulation of cell differentiation. The multiple functions of the IFN system are thought to be an innate defense against microbes and foreign subst ...