微生物学技术

Normal cells in culture divide a defined number of times before reaching replicative senescence (1). The number of divisions that cells can undergo varies with the species of animal and tissue from which the cells are derived. Cells that have exceeded their typical number of divisions are said to h ...

Simian virus 40 large-tumor antigen (SVLT) is a largely nuclear oncogene product (1,2). Evidence from a number of laboratories has indicated that the interaction between cellular nuclear antioncogenes, such as the retinoblastomasusceptibility gene product family (Rb) and p53, med ...

SV40 large T antigen (T-ag) is an oncoprotein that induces transformation of cells through the binding and inactivation of the p53 tumor suppressor protein and the retinoblastoma family of proteins. This chapter focuses on the interaction of T-ag with the product of the retinoblastoma susc ...

Simian virus 40 small t antigen (st) is encoded on a differentially spliced transcript of the SV40 early region. It shares its aminoterminal 82 amino acids with the large T antigen and contains a unique cysteine-rich carboxyterminal half. Small t mutant viruses were obtained originally as dele ...

Simian virus 40 (SV40) infects monkeys and persists in the latent form in the kidneys of this natural host. However, in nonpermissive hosts, such as rodents, the virus induces neoplasia and the outcome is controlled by the immune response of the host (1,2). Virus-neutralizing antibodies are resp ...

Simian virus 40 (SV40) is routinely propagated in established kidney cell lines derived from the African green monkey (1). Generally, one of two cell lines, BSC-1 or CV1, are used for this purpose (2,3). Both of these cell lines are easy to maintain in culture and can be readily frozen and recovered from frozen ...

Several recent investigations using polymerase chain reaction (PCR) detection methods have identified SV40-like DNA sequences in human neoplasms, particularly choroid plexus tumors (1), ependymomas (1), mesotheliomas (2), and osteosarcomas (3,4). Difficulties have arisen ...

The experimental introduction of foreign DNA into the germ line of a mouse is a powerful tool used to identify genetic elements required for appropriate tissue expression, to determine the developmental specificity of gene expression and to determine the consequences of inappropria ...

The human polyomavirus, JCV, is the etiological agent of a fatal central nervous system (CNS) demyelinating disease known as progressive multifocal leukoencephalopathy, or PML. Seroepidemiological studies have indicated that greater than 70% of the human population worldwide is ...

BK virus (BKV), one of two human polyomaviruses, was first isolated in cell culture from the urine of an immunocompromised renal transplant patient in England in 1971 (1). Seroprevalence studies have shown that BK infection is widespread in children throughout the world, and the virus thereaf ...

The Simian virus 40 (SV40) DNA replication system has served as a useful model for studies of DNA synthesis in eukaryotes . One major advantage of this system is that, with the exception of one viral protein, termed T-antigen (T-ag), all of the proteins required for DNA replication are supplied by cells th ...

General properties of human polyomavirus BK (BKV) such as its clinical significance, propagation, and serological assays are described in Chapter 3. In this chapter, two molecular methods for detection of a subtype-specific sequence of BKV are described. Both methods were developed ba ...

The development of soluble cell-free systems that accurately transcribe genes has provided a means to investigate the mechanisms of eukaryotic transcription. The results of many studies have shown that specific DNA cis-acting elements, together with a complex array of protein facto ...

The 5243-bp genome of SV40 contains two transcriptional units (see Fig. 1); the early one, which is first expressed early in the lytic cycle of infection, and the late unit, which is expressed at a significant level only after the onset of viral DNA replication (1). The early genes encode the viral regulato ...

The field of gene therapy has evolved from an investigative curiosity to a major focus of medical research. To date, however, its clinical successes have been few. There are many reasons for this lack of success in clinical studies, but among the most important of these is the inadequacy of gene-delive ...

The use of insect cells and lytic baculovirus for expression of biologically active mammalian proteins has been the method of choice of many investigators. Although prokaryotic expression systems provide higher yields and are technically simpler to use, obtaining biologically ac ...

Many events in the viral life cycle involve protein binding to defined sequences on the viral chromosome. Chromatin immunoprecipitation allows the detection of the in vivo interaction of specific proteins with specific genomic regions. In this technique, living cells are treated with ...

We describe a systematic, high-throughput approach to identify proteins involved in functional adenovirus (Ad)host interactions in vitro and in vivo. We were particularly interested in identifying cellular proteins that interact with fiber knob, which is the moiety within the Ad ca ...

Microinjection of mammalian cells provides a powerful method for analyzing in vivo functions of viral genes and viral gene products. By microinjection, a controlled amount (ranging from several to many thousands of copies) of a viral or cellular gene, a protein product of a gene, a polypeptide f ...

The last 50 yr of molecular biological investigations into human adenoviruses (Ads) have contributed enormously to our understanding of the basic principles of normal and malignant cell growth. Much of this knowledge stems from analyses of the Ad productive infection cycle in permissi ...