Purification Of Microbial β-galactosidase From Kluyveromyces Fragilis By Bioaffinity Partitioning

This work investigated the partitioning of β-galactosidase from Kluyveromyces fragilis in aqueous two-phase systems (ATPS) by bioaffinity. PEG 4000 was chemically activated with thresyl chloride, and the biospecific ligand p-aminophenyl 1-thio-β-D-galactopyranoside (APGP) was attached to the activated PEG 4000. A new two-step method for extraction and purification of the enzyme β-galactosidase from Kluyveromyces fragilis was developed. In the first step, a system composed of 6% PEG 4000-APGP and 8% dextran 505 was used, where β-galactosidase was strongly partitioned to the top phase (K = 2,330). In the second step, a system formed of 13% PEG-APGP and 9% phosphate salt was used to revert the value of the partition coefficient of β-galactosidase (K = 2 x 10-5) in order to provide the purification and recovery of 39% of the enzyme in the bottom salt-rich phase.304324331Aguiñaga-Díaz, P.A., Guzmán, R.Z., Affinity partitioning of metal ions in aqueous polyethylene glycol/salt two-phase systems with PEG-modified dictators (1996) Sep. Sci. Technol., 31 (10), pp. 1483-1499Albertsson, P.Å., (1971) Partition of Cell Particles and Macromolecules. 2nd Ed., p. 323. , New York: InterscienceAlbertsson, P.Å., Tjemeld, F., Phase Diagrams (1994) Meth. Enzumol., 228, pp. 3-13Baskir, J.N., Hatton, T.A., Suter, U.W., Protein partitioning in two-phase aqueous polymer systems (1989) Biotechol. Bioeng., 34 (4), pp. 541-558Birkenmeier, G., Partitioning of blood proteins using immobilized dyes (1994) Meth. 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Population structure, genetic connectivity, and signatures of local adaptation of the giant black tiger shrimp (Penaeus monodon) throughout the indo-pacific region

The giant black tiger shrimp (Penaeus monodon) is native to the Indo-Pacific and is the second most farmed penaeid shrimp species globally. Understanding genetic structure, connectivity, and local adaptation among Indo-Pacific black tiger shrimp populations is important for informing sustainable fisheries management and aquaculture breeding programs. Population genetic and outlier detection analyses were undertaken using 10,593 genome-wide single nucleotide polymorphisms (SNPs) from 16 geographically disparate Indo-Pacific P. monodon populations. Levels of genetic diversity were highest for Southeast Asian populations and were lowest for Western Indian Ocean (WIO) populations. Both neutral (n = 9,930) and outlier (n = 663) loci datasets revealed a pattern of strong genetic structure of P. monodon corresponding with broad geographical regions and clear genetic breaks among samples within regions. Neutral loci revealed seven genetic clusters and the separation of Fiji and WIO clusters from all other clusters, whereas outlier loci revealed six genetic clusters and high genetic differentiation among populations. The neutral loci dataset estimated five migration events that indicated migration to Southeast Asia from the WIO, with partial connectivity to populations in both oceans. We also identified 26 putatively adaptive SNPs that exhibited significant Pearson correlation (P < 0.05) between minor allele frequency and maximum or minimum sea surface temperature. Matched transcriptome contig annotations suggest putatively adaptive SNPs involvement in cellular and metabolic processes, pigmentation, immune response, and currently unknown functions. This study provides novel genome-level insights that have direct implications for P. monodon aquaculture and fishery management practices

Fine-scale population structure and evidence for local adaptation in Australian giant black tiger shrimp (Penaeus monodon) using SNP analysis

Background: Restrictions to gene flow, genetic drift, and divergent selection associated with different environments are significant drivers of genetic differentiation. The black tiger shrimp (Penaeus monodon), is widely distributed throughout the Indian and Pacific Oceans including along the western, northern and eastern coastline of Australia, where it is an important aquaculture and fishery species. Understanding the genetic structure and the influence of environmental factors leading to adaptive differences among populations of this species is important for farm genetic improvement programs and sustainable fisheries management. Results: Based on 278 individuals obtained from seven geographically disparate Australian locations, 10,624 high-quality SNP loci were used to characterize genetic diversity, population structure, genetic connectivity, and adaptive divergence. Significant population structure and differentiation were revealed among wild populations (average FST = 0.001–0.107; p <  0.05). Eighty-nine putatively outlier SNPs were identified to be potentially associated with environmental variables by using both population differentiation (BayeScan and PCAdapt) and environmental association (redundancy analysis and latent factor mixed model) analysis methods. Clear population structure with similar spatial patterns were observed in both neutral and outlier markers with three genetically distinct groups identified (north Queensland, Northern Territory, and Western Australia). Redundancy, partial redundancy, and multiple regression on distance matrices analyses revealed that both geographical distance and environmental factors interact to generate the structure observed across Australian P. monodon populations. Conclusion: This study provides new insights on genetic population structure of Australian P. monodon in the face of environmental changes, which can be used to advance sustainable fisheries management and aquaculture breeding programs

Evolution of the infrared luminosity density and star formation history up to z~1: preliminary results from MIPS

Using deep observations of the Chandra Deep Field South obtained with MIPS at 24mic, we present our preliminary estimates on the evolution of the infrared (IR) luminosity density of the Universe from z=0 to z~1. We find that a pure density evolution of the IR luminosity function is clearly excluded by the data. The characteristic luminosity L_IR* evolves at least by (1+z)^3.5 with lookback time, but our monochromatic approach does not allow us to break the degeneracy between a pure evolution in luminosity or an evolution in both density and luminosity. Our results imply that IR luminous systems (L_IR > 10^11 L_sol) become the dominant population contributing to the comoving IR energy density beyond z~0.5-0.6. The uncertainties affecting our measurements are largely dominated by the poor constraints on the spectral energy distributions that are used to translate the observed 24mic flux into luminosities.Comment: 4 pages, 2 figures. To be published in "Starbursts: From 30 Doradus to Lyman Break Galaxies", held in Cambridge, 6-10 September 2004, Ed. R. de Grijs & R. M. Gonzalez Delgad

Intercalibration of the barrel electromagnetic calorimeter of the CMS experiment at start-up

Calibration of the relative response of the individual channels of the barrel electromagnetic calorimeter of the CMS detector was accomplished, before installation, with cosmic ray muons and test beams. One fourth of the calorimeter was exposed to a beam of high energy electrons and the relative calibration of the channels, the intercalibration, was found to be reproducible to a precision of about 0.3%. Additionally, data were collected with cosmic rays for the entire ECAL barrel during the commissioning phase. By comparing the intercalibration constants obtained with the electron beam data with those from the cosmic ray data, it is demonstrated that the latter provide an intercalibration precision of 1.5% over most of the barrel ECAL. The best intercalibration precision is expected to come from the analysis of events collected in situ during the LHC operation. Using data collected with both electrons and pion beams, several aspects of the intercalibration procedures based on electrons or neutral pions were investigated