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Zhao Q Frederick R Seder K Thao S Sreenath H Peterson F Volkman BF Markley JL Fox BG 《Journal of structural and functional genomics》2004,5(1-2):87-93
The use of 2-L polyethylene terephthalate beverage bottles as a bacterial culture vessel has been recently introduced as an enabling technology for high-throughput structural biology [Sanville Millard, C. et al., 2003. Protein Express. Purif. 29, 311-320]. In the article following this one [Stols et al., this issue, pp. 95-102], this approach was elaborated for selenomethionine labeling used for multiwavelength anomalous dispersion phasing in the X-ray crystallographic determinations of protein structure. Herein, we report an effective and reproducible schedule for uniform 15N- and 13C-labeling of recombinant proteins in 2-L beverage bottles for structural determination by NMR spectroscopy. As an example, three target proteins selected from Arabidopsis thaliana were expressed in Escherichia coli Rosetta (DE3)/pLysS from a T7-based expression vector, purified, and characterized by electrospray ionization mass spectrometry and NMR analysis by 1H-15N heteronuclear single quantum correlation spectroscopy. The results show that expressions in the unlabeled medium provide a suitable control for estimation of the level of production of the labeled protein. Mass spectral characterizations show that the purified proteins contained a level of isotopic incorporation equivalent to the isotopically labeled materials initially present in the growth medium, while NMR analysis of the [U-15N]-labeled proteins provided a convenient method to assess the solution state properties of the target protein prior to production of a more costly double-labeled sample. 相似文献
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Psyllids are insects that harbor endosymbionts (Candidatuus Carsonella ruddii) within specialized cells found in the insect's body cavity. Previous phylogenetic analyses based on endosymbiont
16S–23S ribosomal DNA and a host gene were concordant (M.L. Thao, et al., Appl. Env. Microbiol. 66:2898, 2000). Additional
analyses with atpAGD and rpoBC gave similar trees showing the agreement expected from organisms that evolve through vertical transmission with no gene exchange.
Received: 2 November 2000 / Accepted: 17 November 2000 相似文献
256.
Smith KM Anthony SJ Switzer WM Epstein JH Seimon T Jia H Sanchez MD Huynh TT Galland GG Shapiro SE Sleeman JM McAloose D Stuchin M Amato G Kolokotronis SO Lipkin WI Karesh WB Daszak P Marano N 《PloS one》2012,7(1):e29505
The global trade in wildlife has historically contributed to the emergence and spread of infectious diseases. The United States is the world's largest importer of wildlife and wildlife products, yet minimal pathogen surveillance has precluded assessment of the health risks posed by this practice. This report details the findings of a pilot project to establish surveillance methodology for zoonotic agents in confiscated wildlife products. Initial findings from samples collected at several international airports identified parts originating from nonhuman primate (NHP) and rodent species, including baboon, chimpanzee, mangabey, guenon, green monkey, cane rat and rat. Pathogen screening identified retroviruses (simian foamy virus) and/or herpesviruses (cytomegalovirus and lymphocryptovirus) in the NHP samples. These results are the first demonstration that illegal bushmeat importation into the United States could act as a conduit for pathogen spread, and suggest that implementation of disease surveillance of the wildlife trade will help facilitate prevention of disease emergence. 相似文献
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Lachance N Gareau Y Guiral S Huang Z Isabel E Leclerc JP Léger S Martins E Nadeau C Oballa RM Ouellet SG Powell DA Ramtohul YK Tranmer GK Trinh T Wang H Zhang L 《Bioorganic & medicinal chemistry letters》2012,22(2):980-984
Inhibition of stearoyl-CoA desaturase (SCD) activity represents a potential novel mechanism for the treatment of metabolic disorders including obesity and type II diabetes. To circumvent skin and eye adverse events observed in rodents with systemically-distributed SCD inhibitors, our research efforts have been focused on the search for new and structurally diverse liver-targeted SCD inhibitors. This work has led to the discovery of novel, potent and structurally diverse liver-targeted bispyrrolidine SCD inhibitors. These compounds possess suitable cellular activity and pharmacokinetic properties to inhibit liver SCD activity in a mouse pharmacodynamic model. 相似文献
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Mougenot P Namane C Fett E Camy F Dadji-Faïhun R Langot G Monseau C Onofri B Pacquet F Pascal C Crespin O Ben-Hassine M Ragot JL Van-Pham T Philippo C Chatelain-Egger F Péron P Le Bail JC Guillot E Chamiot-Clerc P Chabanaud MA Pruniaux MP Schmidt F Venier O Nicolaï E Viviani F 《Bioorganic & medicinal chemistry letters》2012,22(7):2497-2502
A novel class of DGAT1 inhibitors containing a thiadiazole core has been discovered. Chemical optimization lead to inhibitors of human DGAT1 with an appropriate ADME profile and that show in vivo activity in target tissues. 相似文献
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