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31.
David M. Alba Salvador Moyà‐Solà Assumpció Malgosa Isaac Casanovas‐Vilar Josep M. Robles Sergio Almécija Jordi Galindo Cheyenn Rotgers Juan Vicente Bertó Mengual 《American journal of physical anthropology》2010,141(1):52-75
Pliopithecus (Pliopithecus) canmatensis sp. nov. is described from several Late Aragonian localities from Abocador de Can Mata (ACM) in els Hostalets de Pierola (Vallès‐Penedès Basin, Catalonia, Spain), spanning from ~11.7 to 11.6 Ma (C5r.3r subchron), and being correlated to the MN8 (reference locality La Grive L3). The ACM remains display a pliopithecine dental morphology with well‐developed pliopithecine triangles on M/2 and M/3. This, together with other occlusal details, negates an attribution to the subgenus Epipliopithecus. Although slightly smaller, the ACM remains are most similar in size to comparable elements of P. piveteaui and P. antiquus. Several occlusal details (such as the greater development of the buccal cingulid in lower molars) and dental proportions (M/3 much longer than M/2), however, indicate greater similarities with P. antiquus from Sansan and La Grive. The ACM remains, however, differ from P. antiquus in dental proportions as well as occlusal morphology of the lower molars (including the less peripheral position of the protoconid and more medial position of the hypoconulid, the more mesial position of the buccal cuspids as compared to the lingual ones, the narrower but distinct mesial fovea, the higher trigonid, and the more extensive buccal cingulid, among others). These differences justify a taxonomic distinction at the species level of the ACM pliopithecid remains with respect to P. antiquus. Previous pliopithecid findings from the Vallès‐Penedès Basin, previously attributed to P. antiquus, are neither attributable to the latter species nor to the newly erected one. Am J Phys Anthropol, 2010. © 2009 Wiley‐Liss, Inc. 相似文献
32.
Hanson MA Brooun A Baker KA Jaakola VP Roth C Chien EY Alexandrov A Velasquez J Davis L Griffith M Moy K Ganser-Pornillos BK Hua Y Kuhn P Ellis S Yeager M Stevens RC 《Protein expression and purification》2007,56(1):85-92
Production of structure-grade mammalian membrane proteins in substantial quantities has been hindered by a lack of methods for effectively profiling multiple constructs expression in higher eukaryotic systems such as insect or mammalian cells. To address this problem, a specialized small-scale eukaryotic expression platform by Thomson Instrument Company (Vertiga-IM) was developed and used in tandem with a Guava EasyCyte microcapillary 96-well cytometer to monitor cell density and health and evaluate membrane protein expression. Two proof of concept experiments were conducted using the human beta(2)-adrenergic receptor (beta(2)AR) and the gap junction protein connexin26 (Cx26) in a baculovirus expression system. First, cell surface expression was used to assess the expression levels of 14 beta(2)AR truncation variants expressed using the Vertiga-IM shaker. Three of these variants were then compared to wild-type beta(2)AR using three metrics: cell surface expression, saturation ligand binding and protein immunoblot analysis of dodecylmaltoside extracted material. Second, a series of systematic Cx26 truncation variants were evaluated for expression by protein immunoblot analysis. The cumulative results for these two systems show that the Vertiga-IM instrument can be used effectively in the parallel insect cell microexpression of membrane protein variants, and that the expression of cell surface molecules as monitored with the Guava EasyCyte instrument can be used to rapidly assess the production of properly folded proteins in the baculovirus expression system. This approach expedites the in vitro evaluation of a large number of mammalian membrane protein variants. 相似文献
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34.
Rapid cultivation of stable aerobic phenol-degrading granules using acetate-fed granules as microbial seed 总被引:10,自引:0,他引:10
cDNA-encoding pyranose 2-oxidase (P2O) from Trametes pubescens was sequenced and cloned into Escherichia coli strain BL21/DE3 on a multicopy plasmid under the control of trc promoter. The synthesis of P2O was studied in a batch culture in M9-based mineral medium: the enzyme was synthesized constitutively at 28 °C in amount corresponding to 8% of the cell soluble protein (0.6 U mg−1). Only small portion of P2O (11%) was in the form of non-active inclusion bodies. Purified recombinant enzyme has similar physico-chemical and kinetic parameters with other P2Os. When compared to the expression of p2o of Trametes ochracea, a ratio of the mature enzyme to inclusion bodies found in the same E. coli host at 28 °C is as much as nine times higher. The finding makes the enzyme from T. pubescens preferable for the large-scale production by recombinant bacteria. The difference in amino acid sequences of the P2O from T. ochracea and T. pubescens may explain the favourable trait of the latter enzyme regarding protein folding. 相似文献
35.
36.
Peti W Page R Moy K O'Neil-Johnson M Wilson IA Stevens RC Wüthrich K 《Journal of structural and functional genomics》2005,6(4):259-267
In structural genomics centers, nuclear magnetic resonance (NMR) screening is in increasing use as a tool to identify folded
proteins that are promising targets for three-dimensional structure determination by X-ray crystallography or NMR spectroscopy.
The use of 1D 1H NMR spectra or 2D [1H,15N]-correlation spectroscopy (COSY) typically requires milligram quantities of unlabeled or isotope-labeled protein, respectively.
Here, we outline ways towards miniaturization of a structural genomics pipeline with NMR screening for folded globular proteins,
using a high-density micro-fermentation device and a microcoil NMR probe. The proteins are micro-expressed in unlabeled or
isotope-labeled media, purified, and then subjected to 1D 1H NMR and/or 2D [1H,15N]-COSY screening. To demonstrate that the miniaturization is functioning effectively, we processed nine mouse homologue protein
targets and compared the results with those from the “macro-scale” Joint Center of Structural Genomics (JCSG) high-throughput
pipeline. The results from the two pipelines were comparable, illustrating that the data were not compromised in the miniaturized
approach. 相似文献
37.
Mathews I Schwarzenbacher R McMullan D Abdubek P Ambing E Axelrod H Biorac T Canaves JM Chiu HJ Deacon AM DiDonato M Elsliger MA Godzik A Grittini C Grzechnik SK Hale J Hampton E Han GW Haugen J Hornsby M Jaroszewski L Klock HE Koesema E Kreusch A Kuhn P Lesley SA Levin I Miller MD Moy K Nigoghossian E Ouyang J Paulsen J Quijano K Reyes R Spraggon G Stevens RC van den Bedem H Velasquez J Vincent J White A Wolf G Xu Q Hodgson KO Wooley J Wilson IA 《Proteins》2005,59(4):869-874
38.
39.
Rife C Schwarzenbacher R McMullan D Abdubek P Ambing E Axelrod H Biorac T Canaves JM Chiu HJ Deacon AM DiDonato M Elsliger MA Godzik A Grittini C Grzechnik SK Hale J Hampton E Han GW Haugen J Hornsby M Jaroszewski L Klock HE Koesema E Kreusch A Kuhn P Lesley SA Miller MD Moy K Nigoghossian E Paulsen J Quijano K Reyes R Sims E Spraggon G Stevens RC van den Bedem H Velasquez J Vincent J White A Wolf G Xu Q Hodgson KO Wooley J Wilson IA 《Proteins》2005,61(2):444-448
40.
Sea urchin spermatozoa are model cells for studying signal transduction events underlying flagellar motility and the acrosome reaction. We previously described the sea urchin sperm receptor for egg jelly 1 (suREJ1) which consists of 1450 amino acids, has one transmembrane segment and binds to the fucose sulfate polymer of egg jelly to induce the sperm acrosome reaction. We also cloned suREJ3 which consists of 2681 amino acids and has 11 putative transmembrane segments. Both these proteins localize to the plasma membrane over the acrosomal vesicle. While cloning suREJ1, we found suREJ2, which consists of 1472 amino acids, has two transmembrane segments and is present in the entire sperm plasma membrane, but is concentrated over the sperm mitochondrion. Experimental evidence suggests that, unlike suREJ1 and suREJ3, suREJ2 does not project extracellularly from the plasma membrane, but is an intracellular plasma membrane protein. All three sea urchin sperm REJ proteins possess a protein module of > 900 amino acids, termed 'the REJ module', that is shared by the human autosomal dominant polycystic kidney disease protein, polycystin-1, and PKDREJ, a testis-specific protein in mammals whose function is unknown. In the present study, we describe the sequence, domain structure and localization of suREJ2 and speculate on its possible function. 相似文献