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Summary Mutations at the URA3 locus of Saccharomyces cerevisiae can be obtained by a positive selection. Wild-type strains of yeast (or ura3 mutant strains containing a plasmid-borne URA3 + gene) are unable to grow on medium containing the pyrimidine analog 5-fluoro-orotic acid, whereas ura3 mutants grow normally. This selection, based on the loss of orotidine-5-phosphate decarboxylase activity seems applicable to a variety of eucaryotic and procaryotic cells.  相似文献   
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Genomic and cDNA clones for three inflorescence-specific genes from Arabidopsis thaliana were isolated and characterized. The genes are tandemly organized in the genome on a 10 kb fragment. The expression of these genes is coordinately regulated in a developmental and organ-specific pattern. They are expressed predominantly in anthers at the later stage of flower development. The primary structure of the encoded gene products exhibits comparable features consisting of a hydrophobic domain at the N-terminal region followed by repeated glycine-rich motifs. Little homology is observed either between the glycine-rich domain of the three genes or with previously described glycine-rich proteins from other plant species.  相似文献   
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Solubilization of fibrin monomers (Fm's) is usually performed with dilute acetic acid, urea or sodium bromide. These solvents can affect the biological properties of Fm's. Therefore we describe a new method to keep Fm's in solution, under milder conditions i.e. by generating them in Dcate solutions and avoiding non-physiological conditions. The in vivo behaviour of iodinated rat Fm's injected in rats and prepared by this new method was compared with that of Fm's dissolved in acetic acid, urea or sodium bromide.Fm's prepared in Dcate solutions accumulate rapidly, within 10 minutes after injection, in all organs tested, predominantly in kidney, liver and lung, probably by interaction with endothelial cells. The blood radioactivity remains nearly constant during the first 90 minutes and decreases thereafter exponentially. Fm's dissolved in sodium bromide behave similarly. However, Fm's dissolved in acetic acid or urea behave differently and do not accumulate in organs. This suggests that Fm's loose their capability to accumulate in organs and probably to interact with endothelial cells when they have been dissolved in acetic acid or urea.The slow exponential clearance phase does not differ significantly between the various Fm's and their T12'S are estimated to lie between 5 and 7 hours.  相似文献   
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Immunoaffinity chromatography with a monoclonal antibody produced against bovine tau protein was used to purify tau proteins from human brain. Fifty grams of brain tissue yielded approximately 2 mg of pure tau proteins. The affinity-purified human tau was used to produce a high-titered rabbit anti-human tau serum. The monoclonal anti-tau antibody and the polyclonal rabbit anti-tau serum were then used to construct a sandwich enzyme-linked immunosorbent assay for detection of human tau proteins, with a sensitivity of 1 ng/ml.  相似文献   
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The multigene family encoding the small subunit polypeptides of ribulose-1,5-bisphosphate carboxylase/oxygenase in the crucifer Arabidopsis thaliana has been isolated and the organization and structure of the individual members determined. The family consists of four genes which have been divided into two subfamilies on the basis of linkage and DNA and amino acid sequence similarities. Three of the genes, designated ats1B, ats2B, and ats3B, reside in tandem on an 8 kb stretch of the chromosome. These genes share greater than 95% similarity in DNA sequence and encode polypeptides identical in length and 96.7% similar in amino acid sequence. The fourth gene, ats1A, is at least 10 kb removed from, or completely unlinked to the B subfamily. The B subfamily genes are more similar to each other than to ats1A in nucleotide and amino acid sequence. All four genes are interupted by two introns whose placement within the coding region of the genes is conserved. The introns of the B subfamily genes are similar in length and nucleotide sequence, but show no similarity to the introns of ats1A. Comparison of the DNA sequences within the immediate 5 and 3 flanking sequences among the genes revealed only limited regions of homology. S1 analysis shows that all four genes are expressed.  相似文献   
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Mighty Piwis defend the germline against genome intruders   总被引:13,自引:0,他引:13  
O'Donnell KA  Boeke JD 《Cell》2007,129(1):37-44
Piwis are a germline-specific subclass of the Argonaute family of RNA interference (RNAi) effector proteins that are associated with a recently discovered group of small RNAs (piRNAs). Recent studies in Drosophila and zebrafish directly implicate Piwi proteins in piRNA biogenesis to maintain transposon silencing in the germline genome (Brennecke et al., 2007; Gunawardane et al., 2007; Houwing et al., 2007). This function may be conserved in mice as loss of Miwi2, a mouse Piwi homolog, leads to germline stem cell and meiotic defects correlated with increased transposon activity (Carmell et al., 2007).  相似文献   
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