101 research outputs found
Expression capable library for studies of Neisseria gonorrhoeae, version 1.0
Background
The sexually transmitted disease, gonorrhea, is a serious health problem in developed as well as in developing countries, for which treatment continues to be a challenge. The recent completion of the genome sequence of the causative agent, Neisseria gonorrhoeae, opens up an entirely new set of approaches for studying this organism and the diseases it causes. Here, we describe the initial phases of the construction of an expression-capable clone set representing the protein-coding ORFs of the gonococcal genome using a recombination-based cloning system.
Results
The clone set thus far includes 1672 of the 2250 predicted ORFs of the N. gonorrhoeae genome, of which 1393 (83%) are sequence-validated. Included in this set are 48 of the 61 ORFs of the gonococcal genetic island of strain MS11, not present in the sequenced genome of strain FA1090. L-arabinose-inducible glutathione-S-transferase (GST)-fusions were constructed from random clones and each was shown to express a fusion protein of the predicted size following induction, demonstrating the use of the recombination cloning system. PCR amplicons of each ORF used in the cloning reactions were spotted onto glass slides to produce DNA microarrays representing 2035 genes of the gonococcal genome. Pilot experiments indicate that these arrays are suitable for the analysis of global gene expression in gonococci.
Conclusion
This archived set of Gateway® entry clones will facilitate high-throughput genomic and proteomic studies of gonococcal genes using a variety of expression and analysis systems. In addition, the DNA arrays produced will allow us to generate gene expression profiles of gonococci grown in a wide variety of conditions. Together, the resources produced in this work will facilitate experiments to dissect the molecular mechanisms of gonococcal pathogenesis on a global scale, and ultimately lead to the determination of the functions of unknown genes in the genome
Recommended from our members
Protein prediction for trait mapping in diverse populations
Genetically regulated gene expression has helped elucidate the biological mechanisms underlying complex traits. Improved high-throughput technology allows similar interrogation of the genetically regulated proteome for understanding complex trait mechanisms. Here, we used the Trans-omics for Precision Medicine (TOPMed) Multi-omics pilot study, which comprises data from Multi-Ethnic Study of Atherosclerosis (MESA), to optimize genetic predictors of the plasma proteome for genetically regulated proteome-wide association studies (PWAS) in diverse populations. We built predictive models for protein abundances using data collected in TOPMed MESA, for which we have measured 1,305 proteins by a SOMAscan assay. We compared predictive models built via elastic net regression to models integrating posterior inclusion probabilities estimated by fine-mapping SNPs prior to elastic net. In order to investigate the transferability of predictive models across ancestries, we built protein prediction models in all four of the TOPMed MESA populations, African American (n = 183), Chinese (n = 71), European (n = 416), and Hispanic/Latino (n = 301), as well as in all populations combined. As expected, fine-mapping produced more significant protein prediction models, especially in African ancestries populations, potentially increasing opportunity for discovery. When we tested our TOPMed MESA models in the independent European INTERVAL study, fine-mapping improved cross-ancestries prediction for some proteins. Using GWAS summary statistics from the Population Architecture using Genomics and Epidemiology (PAGE) study, which comprises ∼50,000 Hispanic/Latinos, African Americans, Asians, Native Hawaiians, and Native Americans, we applied S-PrediXcan to perform PWAS for 28 complex traits. The most protein-trait associations were discovered, colocalized, and replicated in large independent GWAS using proteome prediction model training populations with similar ancestries to PAGE. At current training population sample sizes, performance between baseline and fine-mapped protein prediction models in PWAS was similar, highlighting the utility of elastic net. Our predictive models in diverse populations are publicly available for use in proteome mapping methods at https://doi.org/10.5281/zenodo.4837327
Work and volunteering: longitudinal relationships between work-related experiences and volunteering behaviour
This study examines the effect of work-related experiences on employees’ engagement in community volunteering, using data from a British longitudinal panel of employees. Using a novel analytical approach that separates variation in volunteering due to within-person changes in work conditions from variation due to between-person work differences, we more robustly test existing and new hypotheses about the effects of work on volunteering. New to this literature, we find that commuting and satisfaction with job experiences are significant predictors of community volunteering, both the likelihood to volunteer and volunteering frequency. In turn, volunteering determinants previously explored with cross-sectional data, such as managerial and professional jobs, employment sector and hourly-paid contracts are no longer statistically significant in the within-person models. We discuss a number of important theoretical and practical consequences of these findings
Comparison of Proteomic Assessment Methods in Multiple Cohort Studies
Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/155922/1/pmic13292_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/155922/2/pmic13292.pd
Motivation and satisfaction of volunteers for community-based urban agriculture programmes
Urban agriculture means cultivating plants and raising livestock within cities for food and other uses. A Community‐based Urban Agriculture Programme is where people from residential areas get together as volunteers to practise urban agriculture in an empty space within residential areas. However, the programme encounters problems when it is incapable of attracting enough volunteers and retaining them in order to establish a sustainable programme. This study aims to determine the relationship between the dimensions of motivation and satisfaction of volunteers on the Community‐based Urban Agriculture Programme. Data collected from 375 volunteers on the Community‐based Urban Agriculture Programme in Klang Valley, Malaysia were analysed using descriptive analysis, reliability analysis, correlation analysis, and hierarchical multiple regression analysis. It was found that the most significant predictor of Community‐based Urban Agriculture Programme volunteers’ satisfaction was favoured by external factors such as campaigns, support groups, Department of Extension, and community as well as government policy, followed by love of farming, social referents, and values. Therefore, there should be a focus on the above‐mentioned dimensions of motivation in order to enhance the satisfaction of volunteers towards the Community‐based Urban Agriculture Programme
A comprehensive overview of radioguided surgery using gamma detection probe technology
The concept of radioguided surgery, which was first developed some 60 years ago, involves the use of a radiation detection probe system for the intraoperative detection of radionuclides. The use of gamma detection probe technology in radioguided surgery has tremendously expanded and has evolved into what is now considered an established discipline within the practice of surgery, revolutionizing the surgical management of many malignancies, including breast cancer, melanoma, and colorectal cancer, as well as the surgical management of parathyroid disease. The impact of radioguided surgery on the surgical management of cancer patients includes providing vital and real-time information to the surgeon regarding the location and extent of disease, as well as regarding the assessment of surgical resection margins. Additionally, it has allowed the surgeon to minimize the surgical invasiveness of many diagnostic and therapeutic procedures, while still maintaining maximum benefit to the cancer patient. In the current review, we have attempted to comprehensively evaluate the history, technical aspects, and clinical applications of radioguided surgery using gamma detection probe technology
Corporate Volunteering Climate: Mobilizing Employee Passion for Societal Causes and Inspiring Future Charitable Action
- …