Project description:We undertook gene expression microarray experiments to identify genes that are differentially expressed in different placental and fetal tissue, and resting and Pokeweed Mitogen (PWM) stimulated horse lymphocytes. Conceptus tissues were dissected to obtain chorionic girdle, chorion, and fetal tissue. Freshly isolated horse peripheral blood lymphocytes were split and harvested immediately, or stimulated with PWM and harvested over a five day period. These experiments utilized a custom Agilent horse array designed in house that featured >14,000 probes on an 8x15k array format. Several genes were selected from the results for validation by quantitative real-time PCR. QPCR results matched the microarray results very closely.
Project description:Zooarchaeology by Mass Spectrometry (ZooMS) is a rapidly developing and increasingly utilised peptide mass fingerprinting (PMF) technique that analyses Collagen 1A1 and 1A2 marker peptides for the genus- or species-level identification of fragmentary bones in the archaeological record. Traditionally, this analysis is performed using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-ToF-MS) to identify characteristic m/z values of known marker peptides. Here we present data on the application of a modified ZooMS approach, using nanoflow liquid chromatography – tandem mass spectrometry proteomics, to the analysis of a collection of six early colonial Australian (early to mid-19th Century CE) bone artefacts excavated from a site in Pyrmont, Sydney, Australia in 2017. We were successfully able to identify characteristic marker peptides for bovine COL1A1 and COL1A2 in all six artefacts.
Project description:Mineralised dental plaque (calculus) has proven to be an excellent source of ancient biomolecules. In this study we present a Mycobacterium leprae genome (6.6-fold), the causative agent of leprosy, recovered via shotgun sequencing of 16th century human dental calculus from an individual from Trondheim, Norway. Moreover, ancient mycobacterial peptides were retrieved via mass spectrometry-based proteomics, further validating the presence of the pathogen. M. leprae can readily be detected in the oral cavity and associated mucosal membranes, which likely contributed to it being incorporated into this individual’s dental calculus. This individual showed some possible, but not definitive, evidence of skeletal lesions associated with early stage leprosy. This study is the first known example of successful multi-omics retrieval of M. leprae from archaeological dental calculus. Furthermore, we offer new insights into dental calculus as an alternative sample source to bones or teeth for detecting and molecularly characterizing M. leprae in individuals from the archaeological record.
Project description:We undertook gene expression microarray experiments to identify genes that are differentially expressed in different placental and fetal tissue, and resting and Pokeweed Mitogen (PWM) stimulated horse lymphocytes. Conceptus tissues were dissected to obtain chorionic girdle, chorion, and fetal tissue. Freshly isolated horse peripheral blood lymphocytes were split and harvested immediately, or stimulated with PWM and harvested over a five day period. These experiments utilized a custom Agilent horse array designed in house that featured >14,000 probes on an 8x15k array format. Several genes were selected from the results for validation by quantitative real-time PCR. QPCR results matched the microarray results very closely. Four Day 33-35 chorionic girdle RNAs were compared to matching chorion RNAs, and fetal tissue from two of the conceptuses. Gene expression in resting lymphocytes was compared to gene expression in PWM treated lymphocytes.
Project description:From the 15th to the 19th century, the Trans-Atlantic Slave-Trade influenced the genetic and cultural diversity of numerous populations. We explore genomic and linguistic data from the nine islands of Cabo Verde, the earliest European colony of the era in Africa, a major Slave-Trade platform between the 16th and 19th centuries, and a previously uninhabited location ideal for investigating early admix-ture events between Europeans and Africans. We find that genetic admixture in Cabo Verde occurred primarily between Iberian and Senegambian populations, although forced and voluntary migrations to the archipelago involved numerous other populations. Inter-individual genetic and linguistic variation recapitulate geographic distribution of individuals’ birth-places across Cabo Verdean islands, suggest-ing that Kriolu language variants have developed together with genetic divergences. Furthermore, we find that admixture occurred both early in each island, long before the 18th-century massive TAST deportations triggered by the expansion of plantation economy in Africa and the Americas, and after this era mostly during the abolition of the TAST and of slavery in European colonial empires. Our results illustrate how shifting socio-cultural relationships between enslaved and non-enslaved commu-nities shaped enslaved-African descendants’ genomic diversity and structure on both sides of the At-lantic.