A genomic approach to gene fusion technology

Gene expression profiling provides powerful analyses of transcriptional responses to cellular perturbation. In contrast to DNA array-based methods, reporter gene technology has been underused for this application. Here we describe a genomewide, genome-registered collection of Escherichia coli bioluminescent reporter gene fusions. DNA sequences from plasmid-borne, random fusions of E. coli chromosomal DNA to a Photorhabdus luminescens luxCDABE reporter allowed precise mapping of each fusion. The utility of this collection covering about 30% of the transcriptional units was tested by analyzing individual fusions representative of heat shock, SOS, OxyR, SoxRS, and cya/crp stress-responsive regulons. Each fusion strain responded as anticipated to environmental conditions known to activate the corresponding regulatory circuit. Thus, the collection mirrors E. coli's transcriptional wiring diagram. This genomewide collection of gene fusions provides an independent test of results from other gene expression analyses. Accordingly, a DNA microarray-based analysis of mitomycin C-treated E. coli indicated elevated expression of expected and unanticipated genes. Selected luxCDABE fusions corresponding to these up-regulated genes were used to confirm or contradict the DNA microarray results. The power of partnering gene fusion and DNA microarray technology to discover promoters and define operons was demonstrated when data from both suggested that a cluster of 20 genes encoding production of type I extracellular polysaccharide in E. coli form a single operon.

NO chemical events in the human airway during the immediate and late antigen-induced asthmatic response

A wealth of evidence supports increased NO (NO⋅) in asthma, but its roles are unknown. To investigate how NO participates in inflammatory airway events in asthma, we measured NO⋅ and NO⋅ chemical reaction products [nitrite, nitrate, S-nitrosothiols (SNO), and nitrotyrosine] before, immediately and 48 h after bronchoscopic antigen (Ag) challenge of the peripheral airways in atopic asthmatic individuals and nonatopic healthy controls. Strikingly, NO\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} \begin{equation*}{\mathrm{_{3}^{-}}}\end{equation*}\end{document} was the only NO⋅ derivative to increase during the immediate Ag-induced asthmatic response and continued to increase over 2-fold at 48 h after Ag challenge in contrast to controls [P < 0.05]. NO\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} \begin{equation*}{\mathrm{_{2}^{-}}}\end{equation*}\end{document} was not affected by Ag challenge at 10 min or 48 h after Ag challenge. Although SNO was not detectable in asthmatic airways at baseline or immediately after Ag, SNO increased during the late response to levels found in healthy controls. A model of NO⋅ dynamics derived from the current findings predicts that NO⋅ may have harmful effects through formation of peroxynitrite, but also subserves an antioxidant role by consuming reactive oxygen species during the immediate asthmatic response, whereas nitrosylation during the late asthmatic response generates SNO, safe reservoirs for removal of toxic NO⋅ derivatives.

Stationary-phase mutation in the bacterial chromosome: Recombination protein and DNA polymerase IV dependence

Several microbial systems have been shown to yield advantageous mutations in slowly growing or nongrowing cultures. In one assay system, the stationary-phase mutation mechanism differs from growth-dependent mutation, demonstrating that the two are different processes. This system assays reversion of a lac frameshift allele on an F′ plasmid in Escherichia coli. The stationary-phase mutation mechanism at lac requires recombination proteins of the RecBCD double-strand-break repair system and the inducible error-prone DNA polymerase IV, and the mutations are mostly −1 deletions in small mononucleotide repeats. This mutation mechanism is proposed to occur by DNA polymerase errors made during replication primed by recombinational double-strand-break repair. It has been suggested that this mechanism is confined to the F plasmid. However, the cells that acquire the adaptive mutations show hypermutation of unrelated chromosomal genes, suggesting that chromosomal sites also might experience recombination protein-dependent stationary-phase mutation. Here we test directly whether the stationary-phase mutations in the bacterial chromosome also occur via a recombination protein- and pol IV-dependent mechanism. We describe an assay for chromosomal mutation in cells carrying the F′ lac. We show that the chromosomal mutation is recombination protein- and pol IV-dependent and also is associated with general hypermutation. The data indicate that, at least in these male cells, recombination protein-dependent stationary-phase mutation is a mechanism of general inducible genetic change capable of affecting genes in the bacterial chromosome.

Letters from Benjamin Welles to John Henry Tudor, 1799-1801

Benjamin Welles wrote these six letters to his friend and classmate, John Henry Tudor, between 1799 and 1801. Four of the letters are dated, and the dates of the other two can be deduced from their contents. Welles wrote Tudor four times in September 1799, at the onset of their senior year at Harvard, in an attempt to clear up hurt feelings and false rumors that he believed had caused a chill in their friendship. The cause of the rift is never fully explained, though Welles alludes to "a viper" and "villainous hypocrite" who apparently spread rumors and fueled discord between the two friends. In one letter, Welles asserts that "College is a rascal's Elysium - or the feeling man's hell." In another he writes: "College, Tudor, is a furnace to the phlegmatic, & a Greenland to thee feeling man; it has an atmosphere which breathes contagion to the soul [...] Villains fatten here. College is the embryo of hell." Whatever their discord, the wounds were apparently eventually healed; in a letter written June 26, 1800, Welles writes to ask Tudor about his impending speech at Commencement exercises. In an October 29, 1801 letter, Welles writes to Tudor in Philadelphia (where he appears to have traveled in attempts to recover his failing health) and expresses strong wishes for his friend's recovery and return to Boston. This letter also contains news of their classmate Washington Allston's meeting with painters Henry Fuseli and Benjamin West.

Effects of trace concentrations of anesthetic gases on behavioral performance of operating room personnel /

Mode of access: Internet.

Brief narrative of events touching various reforms /

Last page blank.

Observations on the history and recent proceedings of the Canada Company : addressed in four letters to Frederick Widder, esq., one of the commissioners.

The four letters are dated Hamilton, Gore, 1845 and signed Aliquis.

Hugh Worthington. A novel.

Mode of access: Internet.

Marian Grey ...

Mode of access: Internet.

An arithmetic unit for on-line computation /

Vita.

A four function calculator /

"Report No. UIUCDCS-R-75-692"

Madeline : a novel /

Mode of access: Internet.

L'architetto girovago, opera piacevole ed instruttiva.

Mode of access: Internet.

History of the Church of England : from the abolition of the Roman jurisdiction /

Imprint varies: v.3-4, London, New York, H. Frowde ; Oxford, University Press.--v.5-6, Oxford, Clarendon Press.