Proteomic analysis of rare molecular species of translated polypeptides from a mouse fetal thymus cDNA library

Lefkovits, Ivan; Frey, J. R.

doi:10.1002/1615-9861(200104)1:4<560::aid-prot560>3.0.co;2-s

Cited by 12 publications

(4 citation statements)

References 27 publications

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“…We have acquired knowledge on nucleic acids and protein functions while developing proteomic/genomic approaches that allow the identification of the genetic inheritance and the modulation of gene expression (9)(10)(11). Since there is no definite time point characterizing gene expression, we have chosen 18 h of IFN-γ macrophage activation since proteomic and biological studies are usually done at this time.…”

Section: Discussionmentioning

confidence: 99%

“…Since this library was not normalized it contains several redundant clones. Nucleotide sequences from all the clones were established and are available to the scientific community upon request (9)(10)(11).…”

Section: Analysis Of Gene Expressionmentioning

confidence: 99%

“…The gene profiling technology adopted for the present study (9)(10)(11) using an array constructed from 1536 individual cDNA clones allowed us to study the mentioned defined portion of expressed genes. The choice of the library is of high relevance to the problem under study since in our library there are mainly immunologically relevant gene probes originating from an immunologically active organ, i.e., the fetal thymus.…”

Section: Introductionmentioning

confidence: 99%

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Gene expression in IFN-gamma-activated murine macrophages

Pereira

Modolell

Frey³

et al. 2004

Braz J Med Biol Res

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Macrophages are critical for natural immunity and play a central role in specific acquired immunity. The IFN-γ activation of macrophages derived from A/J or BALB/c mice yielded two different patterns of antiviral state in murine hepatitis virus 3 infection, which were related to a down-regulation of the main virus receptor. Using cDNA hybridization to evaluate mRNA accumulation in the cells, we were able to identify several genes that are differently up-or down-regulated by IFN-γ in A/J (267 and 266 genes, respectively, up-and down-regulated) or BALB/c (297 and 58 genes, respectively, up-and downregulated) mouse macrophages. Macrophages from mice with different genetic backgrounds behave differently at the molecular level and comparison of the patterns of non-activated and IFN-γ-activated A/J or BALB/c mouse macrophages revealed, for instance, an up-regulation and a down-regulation of genes coding for biological functions such as enzymatic reactions, nucleic acid synthesis and transport, protein synthesis, transport and metabolism, cytoskeleton arrangement and extracellular matrix, phagocytosis, resistance and susceptibility to infection and tumors, inflammation, and cell differentiation or activation. The present data are reported in order to facilitate future correlation of proteomic/transcriptomic findings as well as of results obtained from a classical approach for the understanding of biological phenomena. The possible implication of the role of some of the gene products relevant to macrophage biology can now be further scrutinized. In this respect, a down-regulation of the main murine hepatitis virus 3 receptor gene was detected only in IFN-γ-activated macrophages of resistant mice.

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Section: Discussionmentioning

confidence: 99%

Section: Analysis Of Gene Expressionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Gene expression in IFN-gamma-activated murine macrophages

Pereira

Modolell

Frey³

et al. 2004

Braz J Med Biol Res

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“…To achieve this result, each sample is added to a series of different pools, the identity of these subpools providing an “address” of the sample ( 33 ). This process can be visualized by analogy to a 3-D chessboard, where each position represents a sample, and the subpools are the various planes parallel to the top, front, and side: each sample would contribute to three subpools.…”

Section: Identification Of Viral Sourcesmentioning

confidence: 99%

Global Screening for Human Viral Pathogens

Anderson

Gerin

Anderson³

2003

Emerg. Infect. Dis.

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We propose a system for continuing surveillance of viral pathogens circulating in large human populations. We base this system on the physical isolation of viruses from large pooled samples of human serum and plasma (e.g., discarded specimens from diagnostic laboratories), followed by shotgun sequencing of the resulting genomes. The technology for concentrating virions from 100-L volumes was developed previously at Oak Ridge National Laboratory, and the means for purifying and concentrating virions from volumes in microliters have been developed recently. At the same time, marine virologists have developed efficient methods for concentrating, amplifying, and sequencing complex viral mixtures obtained from the ocean. Given this existing technology base, we believe an integrated, automated, and contained system for surveillance of the human “virome” can be implemented within 1 to 2 years. Such a system could monitor the levels of known viruses in human populations, rapidly detect outbreaks, and systematically discover novel or variant human viruses.

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Clonal proteomics: One gene - family of proteins

Kettman¹,

Coleclough

Frey³

et al. 2002

Proteomics

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The work presented here attempts to consolidate our knowledge on cellular transcriptome and proteome. It takes into account that a typical activated cell (lymphocyte) contains 40 000 mRNA molecules at any time, and it represents about 5000 different molecular species of transcripts. Such a cell has about 1 000 000 000 protein molecules, some of them being present at 10 000 000 copies while others at a very low copy number (say 1 to 10 copies per cell). By studying cell free expression of individual cDNA clones (or pools of known complexity) we address to those rare molecular components that will remain undetected by the current analytical means. For our analysis we use cell free translation systems (wheat germ or rabbit reticulocyte origin) and we study polypeptide products originating from intact, or restriction endonuclease-treated cDNA clones. We conclude that in most instances expressed genes yield transcript(s) that translate into several, and often very numerous families of polypeptide species. In our ISODALT two-dimensional gel system we characterize the proteomic profile of the clonal polypeptide families in terms of their molecular mass, charge, multiple products, and appearance.

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Proteomic analysis of rare molecular species of translated polypeptides from a mouse fetal thymus cDNA library

Cited by 12 publications

References 27 publications

Gene expression in IFN-gamma-activated murine macrophages

Gene expression in IFN-gamma-activated murine macrophages

Global Screening for Human Viral Pathogens

Clonal proteomics: One gene - family of proteins

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