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51.
EP Guthrie T Quinton-Jager LS Moran BE Slatko RB Kucera JS Benner GG Wilson JE Brooks 《Canadian Metallurgical Quarterly》1996,180(1-2):107-112
SphI, a type II restriction-modification (R-M) system from the bacterium Streptomyces phaeochromogenes, recognizes the sequence 5'-GCATGC. The SphI methyltransferase (MTase)-encoding gene, sphIM, was cloned into Escherichia coli using MTase selection to isolate the clone. However, none of these clones contained the restriction endonuclease (ENase) gene. Repeated attempts to clone the complete ENase gene along with sphIM in one step failed, presumably due to expression of SphI ENase gene, sphIR, in the presence of inadequate expression of sphIM. The complete sphIR was finally cloned using a two-step process. PCR was used to isolate the 3' end of sphIR from a library. The intact sphIR, reconstructed under control of an inducible promoter, was introduced into an E. coli strain containing a plasmid with the NlaIII MTase-encoding gene (nlaIIIM). The nucleotide sequence of the SphI system was determined, analyzed and compared to previously sequenced R-M systems. The sequence was also examined for features which would help explain why sphIR unlike other actinomycete ENase genes seemed to be expressed in E. coli. 相似文献
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J Ali EB Freyfogle RJ Parker RM Bell C Maimaris BE Krantz IS Hughes 《Canadian Metallurgical Quarterly》1997,163(7):483-486
OBJECTIVES: To determine whether blurred vision caused by exposure to triethylamine (TEA) can be detected by the measurement of contrast sensitivity. METHODS: 41 cold box core makers of three foundries and 82 control workers were examined. A detailed ocular and medical history was obtained from the subjects. The contrast sensitivity of the core makers was measured on Monday and Friday of the same week both before and immediately after work and also on a third day, when air samples of TEA were collected. Contrast sensitivity and visual acuity were measured by optotype figures at full contrast, 2.5% contrast, and 0.6% contrast. The changes in contrast sensitivity were used for the analysis. The results of binocular vision and the results of the dominant eye were analysed. Urine specimens for the analysis of TEA were collected on every occasion when contrast sensitivity was measured. RESULTS: 78% of the core makers had had symptoms of blurred vision, and 31% had had trouble driving or working. The breathing zone eight hour time weighted average TEA concentrations were 0.3-60 mg/m3. The mean urinary TEA concentration after the shift was 35 mmol/mol creatinine. Continuous monitoring showed high peaks of TEA leakage at a core making machine. Changes in binocular visual acuity did not differ between the exposed and unexposed workers. The contrast sensitivity decreased in 49% of the core makers and 21% of the controls (P = 0.002). CONCLUSIONS: The blurred vision caused by exposure to TEA can be documented by measuring contrast sensitivity. The mechanism by which TEA produces symptoms remains an issue of further study. 相似文献
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Lymphocytes of a New World primate, the cotton-top tamarin (Saguinus oedipus), express classical G-related major histocompatibility complex (MHC) class I molecules with unusually limited polymorphism and variability. Three G-related loci, an F locus, an E locus, and two pseudogenes (So-N1 and So-N3) have been identified by cDNA library screening and extensive PCR analysis of both cDNA and genomic DNA from the cotton-top tamarin. Furthermore, each genus of the subfamily Callitrichinae (tamarins and marmosets) appears to express its own unique set of MHC class I genes, likely due to a rapid turnover of loci. The rapid emergence of unique MHC class I genes in the Callitrichinae genera, resulting from an active process of duplication and inactivation of loci, may account for the limited diversity of the MHC class I genes in the cotton-top tamarin. To determine the nature of the entire complement of MHC class I genes in the cotton-top tamarin, we synthesized a genomic DNA library and screened it with MHC class I-specific probes. We isolated nine new MHC class I pseudogenes from this library. These newly isolated tamarin G-related MHC class I pseudogenes are not closely related to any of their functional counterparts in the tamarin, suggesting that they do not share a recent common ancestral gene with the tamarin's currently expressed MHC class I loci. In addition, these tamarin sequences display a high rate of nonsynonymous substitutions in their putative peptide binding region. This indicates that the genes from which they have derived were likely subject to positive selection and, therefore, were once functional. Our data support the notion that an extremely high rate of loci turnover is largely responsible for the limited diversity of the MHC class I genes in the cotton-top tamarin. 相似文献
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Escherichia coli O29:H21 is a human enterotoxigenic serotype that produces heat-stable (ST-I) enterotoxin, adheres diffusely to HeLa cells, and presents colonization factor antigen IV (CFA/IV) composed of CS5CS6 surface antigens. In one strain studied the genes for diffuse adherence and CFA/IV (CS5CS6) production were found to be present in the same plasmid encoding ST-I. The virulence plasmid (Ent) presented two unrelated basic replicons homologous to repFIC and repW. Gene(s) encoding diffuse adherence did not share homology with the probe for F1845 fimbrial adhesin which is responsible for this phenotype in other E. coli strains. Ent plasmids containing genes for diffuse adherence have not been described previously. 相似文献
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