Nature of {varphi}X174 Linear DNA from a DNA Ligase-Defective Host

Linear DNAs have been prepared from {varphi}X phage and from {varphi}X RF II (double-stranded circular form of {varphi}X DNA, formed during infection and nicked in one or both strands) molecules derived from infection at the restrictive temperature of Escherichia coli ts7, a host mutant with a temperature-sensitive DNA ligase activity. The linear DNA from these phages can be circularized by annealing with fragments of {varphi}X RF DNA produced by the Haemophilus influenzae restriction nuclease. The circularization experiment indicated that the site of breakage of the linear phage DNAs is not unique nor confined to a particular region of the genome. These linear DNAs were less than 0.1% as infective as circular phage DNA. The linear, positive strand of late RF II DNA, however, is uniquely nicked in the region of the {varphi}X genome corresponding to cistron A. Although a low level of infectivity is associated with the linear DNA derived from late RF II, this infectivity appears to be a result of the association of linear positive and linear negative strands during the infectivity assay

Discovering and Assessing Desired Student Financial Services at East Tennessee State University.

The purpose of this study was to discover and assess student financial services delivered to students enrolled at East Tennessee State University. The research was undertaken for institutional self-improvement. The research explored changes that have occurred in student financial services in the dynamic higher education market. The research revealed universities pursued best practices for the delivery of student financial services through expanded employee knowledge, restructured organizations, and integrated information technologies. The research was conducted during October and November, 2006. The data were gathered from an online student survey of student financial services. The areas researched included: the Bursar office, the Financial Aid office, and online services. The results of the data analysis revealed problems with the students\u27 perceived quality of existing financial services and the additional services students desire. The research focused on student perceptions of the quality of financial services by age and gender classifications and response categories. Although no statistically significant difference was found between the age-gender classifications on the perception of the quality of the financial services studied, the research adds to our understanding of student financial services at East Tennessee State University. Recommendation for continued research included annual surveys of segmented student populations that include ethnicity, age, gender, and educational level. The research would be used for continuous improvement efforts and student relationship management. Also additional research was recommended for employee learning in relation to the institution\u27s mission, goals, and values

Control of Baculovirus Polyhedrin Gene Expression by Very Late Factor 1

Abstractvlf-1is a baculovirus gene that regulates very late gene expression (J. R. McLachlin and L. K. Miller,J. Virol.,68, 7746–7756, 1994) and also plays a crucial role in the replication of the budded form ofAutographa californicanuclear polyhedrosis virus (AcMNPV) (S. Yang and L. K. Miller, “Expression and mutational analysis of the baculovirus very late factor 1 (vlf-1) gene.”Virology,245, 99–109, 1998). To examine the influence ofvlf-1expression on baculovirus infection, we constructed recombinant viruses that expressed only low levels of VLF-1 and recombinants withvlf-1under the control of different promoters. Viruses with mutant alleles ofvlf-1that produced low levels of VLF-1 replicated the budded form of the virus normally but produced no occlusion bodies. Thus, a higher concentration of VLF-1 was needed to activate very late gene expression than was needed to support budded virus production. By altering the level and/or timing ofvlf-1expression, the timing of polyhedrin gene (polh) expression, which normally occurs very late in infection, could be advanced or delayed. Early overexpression ofvlf-1increased the level of expression from thepolhpromoter but caused premature cellular disintegration. The data indicate that VLF-1 is the limiting factor in very late gene expression and that the level of VLF-1 controls the onset of occlusion

Expression and functional analysis of a baculovirus gene encodinga truncated protein kinase homolog

AbstractAutographa californica nuclear polyhedrosis virus (AcMNPV) potentially encodes a 215-amino acid polypeptide containing6 out of 11 motifs conserved among eukaryotic protein kinases (Morris et al., Virology 200, 360–369, 1994). We examined the expression of this gene, named pk2, at the transcriptional and translational levels and the possible role of the gene during baculovirus replication in cell culture and insect larvae. Northern (RNA) blot analysis revealed that pk2 was transcribed primarily as an early 1.2-kb RNA. Western blot analysis showed that pk2 was expressed as a 25-kDa protein, PK2, which was present both early and late during virus infection. To examine the function(s) of pk2, we constructed a mutant baculovirus, vKINdel, in which one-third of the PK2-coding region was deleted and then compared the characteristics of vKINdel with wild-type AcMNPV and a marker-rescued revertant. The pk2 deletion mutation had no discernable effect on the number, size, or appearance of plaques, the kinetics of protein synthesis or protein phosphorylation profiles during virus infection of cultured SF-21 cells. Deletion of pk2 also had no significant influence on the infectivity or virulence of the baculovirus in larval bioassays and the level of occluded virus production was normal. Thus, pk2 does not appear to have a significant influence on virus replication in the host systems examined

Harvest-based Monitoring in the Inuvialuit Settlement Region: Steps for Success

We define harvest-based monitoring as the long-term collection of data or samples from a subsistence harvest in order to reveal, document, and track changes in biophysical resources. Our objective is to describe five practical steps that have guided us over the past two decades during delivery of harvest-based monitoring studies in the Inuvialuit Settlement Region (ISR). Studies have usually been designed to detect (but not necessarily explain) change, to involve local harvesters, and to incorporate indigenous and science-based knowledge. The five steps are to (1) formulate a scientific research or long-term monitoring question that can reasonably be answered by analyzing data from harvests or harvested specimens, (2) design the program according to scientific and indigenous protocols, (3) determine respective partner roles for delivery of the field program, (4) conduct the field work, and (5) analyze data and communicate results. At all steps, it is important to ensure that science and indigenous knowledge partners respect and trust each other’s skills, knowledge, and abilities; that regular communication is fostered; and that provisions are in place to monitor progress. The credible blending of indigenous and scientific views and skills improves the likelihood of ultimately understanding the resource, its habitats, and its inherent ecological relationships.Key words: harvest-based monitoring, Inuvialuit Settlement Region, collaborative research, participatory researchNous définissons la surveillance des captures comme la collecte à long terme de données ou d’échantillons provenant des captures ou récoltes de subsistance et ce, dans le but de révéler, de documenter et de suivre les changements caractérisant les ressources biophysiques. Notre objectif consiste à décrire cinq étapes pratiques qui nous ont servi de guides ces deux dernières décennies dans le cadre d’études de surveillance des captures dans la région désignée des Inuvialuit (RDI). Habituellement, les études sont conçues pour détecter (et non pas nécessairement pour expliquer) le changement, pour faire appel aux personnes faisant les captures dans la région et pour favoriser l’intégration des connaissances indigènes et scienti­fiques. Ces cinq étapes sont les suivantes : 1) formuler une question de recherche scientifique ou de surveillance à long terme à laquelle on peut raisonnablement répondre au moyen de l’analyse des données de captures ou des échantillons capturés; 2) concevoir un programme qui respecte les protocoles scientifiques et indigènes; 3) déterminer le rôle des partenaires respectifs en ce qui a trait à l’exécution du programme sur le terrain; 4) réaliser le travail sur le terrain; et 5) analyser les données puis communiquer les résultats. À toutes ces étapes, il est important de faire en sorte que les partenaires en matière de connaissances scientifiques et de connaissances indigènes respectent les compétences, les connaissances et les aptitudes de chacun, et se fassent confiance; que les partenaires communiquent régulièrement; et que des dispositions soient en place pour suivre les progrès. Le mélange crédible de points de vue et de compétences indigènes et scientifiques améliore la probabilité que l’on finisse par comprendre la ressource, ses habitats et ses relations écologiques inhérentes. Mots clés : surveillance des captures, région désignée des Inuvialuit, recherche collaborative, recherche participativ

A Preliminary List of the Lichens of Dickinson and Emmet Counties, lowa

A list of lichen species collected in Dickinson and Emmet Counties is presented

A Baculovirus Mutant Defective in PKIP, a Protein Which Interacts with a Virus-Encoded Protein Kinase

AbstractWe have found that a temperature-sensitive mutant of the baculovirus AcMNPV, tsB97, is defective in PKIP, the product of ORF24 which was previously found to interact with and stimulate the activity of a virus-encoded protein kinase, PK-1. The mutant lacks the ability to form plaques and occlusion bodies at the nonpermissive temperature. The mutant displays several properties which suggest a defect in the latter half of the late phase of infection; these properties include a delay in the shutoff of host protein synthesis, the presence of aberrant electron-dense bodies associated with the virogenic stroma, and the production of few, if any, progeny budded virus. A study of the expression of selected late genes showed no difference in the timing or level of transcription or translation of most late genes. However, elevated levels of the late 6.9K protein, a protamine-like protein, were observed in mutant-infected cells at 24 h postinfection, suggesting a defect in the regulation of this protein. Two polypeptides, 40 and 6 kDa, exhibited considerably higher levels of steady-state phosphorylation in wt-infected cells versus tsB97-infected cells at 24 h p.i. and could be candidates for PK-1/PKIP-mediated phosphorylation. The tsB97 mutant also displayed a severe defect in very late gene transcription which accounts for its inability to form occlusion bodies. The effect of PKIP on very late gene transcription may be a secondary effect of the block in the late phase of infection. PKIP showed no ability to transactivate expression from a very late promoter in transient expression assays

Wind Signatures In The X-Ray Emission-Line Profiles Of The Late-O Supergiant Zeta Orionis

X-ray line-profile analysis has proved to be the most direct diagnostic of the kinematics and spatial distribution of the very hot plasma around O stars. The Doppler-broadened line profiles provide information about the velocity distribution of the hot plasma, while the wavelength-dependent attenuation across a line profile provides information about the absorption to the hot plasma, thus providing a strong constraint on its physical location. In this paper, we apply several analysis techniques to the emission lines in the Chandra High Energy Transmission Grating Spectrometer (HETGS) spectrum of the late-O supergiant zeta Ori (O9.7 Ib), including the fitting of a simple line-profile model. We show that there is distinct evidence for blueshifts and profile asymmetry, as well as broadening in the X-ray emission lines of zeta Ori. These are the observational hallmarks of a wind-shock X-ray source, and the results for zeta Ori are very similar to those for the earlier O star, zeta Pup, which we have previously shown to be well fit by the same wind-shock line-profile model. The more subtle effects on the line-profile morphologies in zeta Ori, as compared to zeta Pup, are consistent with the somewhat lower density wind in this later O supergiant. In both stars, the wind optical depths required to explain the mildly asymmetric X-ray line profiles imply reductions in the effective opacity of nearly an order of magnitude, which may be explained by some combination of mass-loss rate reduction and large-scale clumping, with its associated porosity-based effects on radiation transfer. In the context of the recent reanalysis of the helium-like line intensity ratios in both zeta Ori and zeta Pup, and also in light of recent work questioning the published mass-loss rates in OB stars, these new results indicate that the X-ray emission from zeta Ori can be understood within the framework of the standard wind-shock scenario for hot stars