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( Wen Fei Xue ),( Jing Fu Peng ),( Xiao Li Yu ),( Shu Lin Zhang ),( Boping Zhou ),( Dan Qing Jiang ),( Jian Bo Chen ),( Bing Bing Ding ),( Bin Zhu ),( Yao Li ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.1
The widespread occurrence of drug-resistant Mycobacterium tuberculosis places importance on the detection of TB (tuberculosis) drug susceptibility. Conventional drug susceptibility testing (DST) is a lengthy process. We developed a rapid enzymatic color-reaction-based biochip assay. The process included asymmetric multiplex PCR/templex PCR, biochip hybridization, and an enzymatic color reaction, with specific software for data operating. Templex PCR (tem- PCR) was applied to avoid interference between different primers in conventional multiplex- PCR. We applied this assay to 276 clinical specimens (including 27 sputum, 4 alveolar lavage fluid, 2 pleural effusion, and 243 culture isolate specimens; 40 of the 276 were non-tuberculosis mycobacteria specimens and 236 were M. tuberculosis specimens). The testing process took 4.5 h. A sensitivity of 50 copies per PCR was achieved, while the sensitivity was 500 copies per PCR when tem-PCR was used. Allele sequences could be detected in mixed samples at aproportion of 10%. Detection results showed a concordance rate of 97.46% (230/236) in rifampicin resistance detection (sensitivity 95.40%, specificity 98.66%) and 96.19% (227/236) in isoniazid (sensitivity 93.59%, specificity 97.47%) detection with those of DST assay. Concordance rates of testing results for sputum, alveolar lavage fluid, and pleural effusion specimens were 100%. The assay provides a potential choice for TB diagnosis and treatment.
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Dandan BIAN,Xiaoming ZHAO,Li CHEN,Jiwu TIAN,Qiuning LIU,Chunlin ZHOU,Boping Tang 한국곤충학회 2018 Entomological Research Vol.48 No.1
Eukaryotic initiation factor 5A (eIF‐5A) is a highly conserved protein found in all eukaryotic organisms that plays a key role in the regulation of many cellular processes including translation elongation, cell proliferation, programmed cell death, mRNA turnover and decay, and abiotic stress responses. In this study, the eIF‐5A gene from the Chinese oak silkworm Antheraea pernyi (Lepidoptera: Saturniidae) was characterized. The full‐length ApeIF‐5A cDNA of 1056 bp includes a 5′‐untranslated region (UTR) of 138 bp, a 3′‐UTR of 435 bp, and an open reading frame of 483 bp encoding a polypeptide of 160 amino acids. The deduced ApeIF‐5A protein shares 99 %, 82 %, and 72 % sequence identity with orthologs in Bombyx mori, Drosophila melanogaster and Homo sapiens, indicating high conservation during animal evolution. Real‐time quantitative reverse transcription PCR revealed expression in all four developmental stages and in all nine tissues tested, consistent with an important role in development. After challenge with lipopolysaccharide, the expression levels of ApeIF‐5A were markedly upregulated. Phylogenetic analysis of amino acid sequences revealed A. pernyi eIF‐5A was closely related to B. mori eIF‐5A, consistent with traditional classification and other molecular data. The results indicate the potential value of eIF‐5A in phylogenetic analysis.
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