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排序方式: 共有192条查询结果,搜索用时 15 毫秒
81.
Sathiya N. Manivannan Thomas L. Jacobsen Peter Lyon Bhavani Selvaraj Peter Halpin Amanda Simcox 《Genetics》2015,201(4):1319-1328
Transfection of transgenes into Drosophila cultured cells is a standard approach for studying gene function. However, the number of transgenes present in the cell following transient transfection or stable random integration varies, and the resulting differences in expression level affect interpretation. Here we developed a system for Drosophila cell lines that allows selection of cells with a single-copy transgene inserted at a specific genomic site using recombination-mediated cassette exchange (RMCE). We used the φC31 integrase and its target sites attP and attB for RMCE. Cell lines with an attP-flanked genomic cassette were transfected with donor plasmids containing a transgene of interest (UAS-x), a dihydrofolate reductase (UAS-DHFR) gene flanked by attB sequences, and a thymidine kinase (UAS-TK) gene in the plasmid backbone outside the attB sequences. In cells undergoing RMCE, UAS-x and UAS-DHFR were exchanged for the attP-flanked genomic cassette, and UAS-TK was excluded. These cells were selected using methotrexate, which requires DHFR expression, and ganciclovir, which causes death in cells expressing TK. Pure populations of cells with one copy of a stably integrated transgene were efficiently selected by cloning or mass culture in ∼6 weeks. Our results show that RMCE avoids the problems associated with current methods, where transgene number is not controlled, and facilitates the rapid generation of Drosophila cell lines in which expression from a single transgene can be studied. 相似文献
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83.
Justin Bahl Scott Krauss Denise Kühnert Mathieu Fourment Garnet Raven S. Paul Pryor Lawrence J. Niles Angela Danner David Walker Ian H. Mendenhall Yvonne C. F. Su Vivien G. Dugan Rebecca A. Halpin Timothy B. Stockwell Richard J. Webby David E. Wentworth Alexei J. Drummond Gavin J. D. Smith Robert G. Webster 《PLoS pathogens》2013,9(8)
Wild birds have been implicated in the emergence of human and livestock influenza. The successful prediction of viral spread and disease emergence, as well as formulation of preparedness plans have been hampered by a critical lack of knowledge of viral movements between different host populations. The patterns of viral spread and subsequent risk posed by wild bird viruses therefore remain unpredictable. Here we analyze genomic data, including 287 newly sequenced avian influenza A virus (AIV) samples isolated over a 34-year period of continuous systematic surveillance of North American migratory birds. We use a Bayesian statistical framework to test hypotheses of viral migration, population structure and patterns of genetic reassortment. Our results reveal that despite the high prevalence of Charadriiformes infected in Delaware Bay this host population does not appear to significantly contribute to the North American AIV diversity sampled in Anseriformes. In contrast, influenza viruses sampled from Anseriformes in Alberta are representative of the AIV diversity circulating in North American Anseriformes. While AIV may be restricted to specific migratory flyways over short time frames, our large-scale analysis showed that the long-term persistence of AIV was independent of bird flyways with migration between populations throughout North America. Analysis of long-term surveillance data provides vital insights to develop appropriately informed predictive models critical for pandemic preparedness and livestock protection. 相似文献
84.
Y Deng J Zhao D Sakurai KM Kaufman JC Edberg RP Kimberly DL Kamen GS Gilkeson CO Jacob RH Scofield CD Langefeld JA Kelly ME Alarcón-Riquelme BIOLUPUS GENLES Networks JB Harley TJ Vyse BI Freedman PM Gaffney KM Sivils JA James TB Niewold RM Cantor W Chen BH Hahn EE Brown PROFILE BP Tsao 《Arthritis research & therapy》2012,14(Z3):A5
85.
Ulf-Peter Apfel Christian R. Kowol Yvonne Halpin Florian Kloss Joachim Kübel Helmar Grls Johannes G. Vos Bernhard K. Keppler Enrico Morera Gino Lucente Wolfgang Weigand 《Journal of inorganic biochemistry》2009,103(9):1236-1244
The present investigations deal with the modeling of the peptide surrounding of [FeFe] hydrogenase using amine containing disulphides to simulate possible influences of the amino acid lysine (K237) on the electrochemical and electrocatalytic properties of biomimetic compounds based on [Fe2S2] moieties. Fe3(CO)12 was reacted with Boc-4-amino-1,2-dithiolane, Boc-Adt-OMe (Adt = 4-amino-1,2-dithiolane-4-carboxylic acid, Boc = tert-butoxycarbonyl) and Boc-Adp tert-butyl ester (Adp = (S)-2-amino-3-(1,2-dithiolan-4-yl)propionic acid) to elongate the FeN distance in comparison to the well known [Fe2{(SCH2)2NR}(CO)6] model complexes. Efforts to deprotect the complexes containing Boc-4-amino-1,2-dithiolane with trifluoroacetic acid result in the formation of [Fe3(μ3-O)(μ-O2C2F3)6(OC4H8)2(H2O)]. The novel [2Fe2S] complexes are characterized using spectroscopic, electrochemical techniques and X-ray diffraction studies. 相似文献
86.
E M Rosen I D Goldberg H M Shapiro S E Levenson P A Halpin D Faraggi 《Journal of cellular physiology》1986,128(2):337-344
Smooth muscle cell (SMC) growth may play an important role in the pathogenesis of vascular diseases such as atherosclerosis and hypertension. Recent studies have demonstrated that, under different growth stimuli in vivo, SMC may respond by proliferation of diploid cells, polyploidization to the tetraploid (or even octaploid) state, or both. In this study, we used flow cytometry to evaluate the intrinsic tendencies of aortic SMC and nonarterial cells from rats of different strains, ages, and blood pressures to polyploidize in response to in vitro growth stimulation. Significant strain-related differences in polyploidization of aortic SMC were found (P less than 0.001): highest in WKY (normotensive inbred rat related to SHR), intermediate in SHR (genetically hypertensive rat), and lowest in Sprague-Dawley and Fischer (normotensive outbred and inbred rats). Animal age had less or no effect on the degree of polyploidization. Nonarterial cells (venous SMC and lung cells) from WKY and SHR remained essentially diploid, suggesting tissue specificity of in vitro polyploidization. Studies of the growth kinetics of uncloned and clonal populations of aortic SMC revealed decreased proliferation as the ploidy increased in WKY, SHR, and Sprague-Dawley. These findings suggest that genetic strain factors as well as cell type/site of origin significantly influence in vitro polyploidization, whereas animal age and blood pressure do not. The findings also emphasize the need to consider ploidy changes when evaluating in vitro SMC growth kinetics. Further studies will improve understanding of SMC growth regulation and the functional significance of vascular polyploidy. 相似文献
87.
88.
Identification and characterisation of cDNA clones encoding cinnamyl alcohol dehydrogenase from tobacco 总被引:15,自引:0,他引:15
Cinnamyl alcohol dehydrogenase (CAD, EC 1.1.1.195) is an enzyme involved in lignin biosynthesis. We have previously isolated pure CAD enzyme as two closely related polypeptides of 44 and 42.5 kDa from tobacco stems. In this paper, we report partial amino acid sequences of these two polypeptides. Based on the peptide sequences mixed oligonucleotides were used to screen a tobacco stem cDNA library and CAD cDNA clones encoding the two polypeptides were identified. DNA sequence comparisons indicate very high sequence identity between these clones both in the coding and in the 5 and 3 untranslated sequences. The close similarity between the two CAD genes leads us to suggest that they do not represent different isoforms but are the same gene from each of the two parental lines of Nicotiana tabacum cv. Samsun. Sequence comparisons with alcohol dehydrogenase 1 (ADH1) from yeast shows sequence similarities of ca. 30%, while comparisons with maize, barley and potato ADH1 sequences show similarities of not more than 23%.Abbreviations CAD
cinnamyl alcohol dehydrogenase (EC 1.1.1.195)
- ADH
alcohol dehydrogenase (EC 1.1.1.1) 相似文献
89.
Models of rheumatoid arthritis (RA) in laboratory animals are important tools for research into pathogenic mechanisms and
the development of effective, safe therapies. Rodent models (rats and mice) have provided important information about the
pathogenic mechanisms. However, the evolutionary distance between rodents and humans hampers the translation of scientific
principles into effective therapies. The impact of the genetic distance between the species is especially seen with treatments
based on biological molecules, which are usually species-specific. The outbred nature and the closer anatomical, genetic,
microbiological, physiological, and immunological similarity of nonhuman primates to humans may help to bridge the wide gap
between inbred rodent strain models and the heterogeneous RA patient population. Here we review clinical, immunological and
pathological aspects of the rhesus monkey model of collagen-induced arthritis, which has emerged as a reproducible model of
human RA in nonhuman primates. 相似文献
90.