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White, H,Deprez, L,Corbisier, P,Hall, V,Lin, F,Mazoua, S,Trapmann, S,Aggerholm, A,Andrikovics, H,Akiki, S,Barbany, G,Boeckx, N,Bench, A,Catherwood, M,Cayuela, J-M,Chudleigh, S,Clench, T,Colomer, D,Dar Nature Publishing Group 2015 Leukemia Vol.29 No.2
<P>Serial quantification of <I>BCR–ABL1</I> mRNA is an important therapeutic indicator in chronic myeloid leukaemia, but there is a substantial variation in results reported by different laboratories. To improve comparability, an internationally accepted plasmid certified reference material (CRM) was developed according to ISO Guide 34:2009. Fragments of <I>BCR–ABL1</I> (e14a2 mRNA fusion)<I>, BCR</I> and <I>GUSB</I> transcripts were amplified and cloned into pUC18 to yield plasmid pIRMM0099. Six different linearised plasmid solutions were produced with the following copy number concentrations, assigned by digital PCR, and expanded uncertainties: 1.08±0.13 × 10<SUP>6</SUP>, 1.08±0.11 × 10<SUP>5</SUP>, 1.03±0.10 × 10<SUP>4</SUP>, 1.02±0.09 × 10<SUP>3</SUP>, 1.04±0.10 × 10<SUP>2</SUP> and 10.0±1.5 copies/μl. The certification of the material for the number of specific DNA fragments per plasmid, copy number concentration of the plasmid solutions and the assessment of inter-unit heterogeneity and stability were performed according to ISO Guide 35:2006. Two suitability studies performed by 63 <I>BCR–ABL1</I> testing laboratories demonstrated that this set of 6 plasmid CRMs can help to standardise a number of measured transcripts of e14a2 <I>BCR–ABL1</I> and three control genes (<I>ABL1, BCR</I> and <I>GUSB</I>). The set of six plasmid CRMs is distributed worldwide by the Institute for Reference Materials and Measurements (Belgium) and its authorised distributors (https://ec.europa.eu/jrc/en/reference-materials/catalogue/; CRM code ERM-AD623a-f).</P>
Artymovich, K.,Kim, J.S.,Linz, J.E.,Hall, D.F.,Kelley, L.E.,Kalbach, H.L.,Kathariou, S.,Gaymer, J.,Paschke, B. Academic Press 2013 Food microbiology Vol.34 No.2
Campylobacter jejuni is an important foodborne pathogen of humans and its primary reservoir is the gastrointestinal (GI) tract of chickens. Our previous studies demonstrated that phase variation to specific ''successful alleles'' at C. jejuni contingency loci Cj0045 (successful alleles carry 9G or 10G homopolymeric tracts) and Cj0170 (successful allele carries a 10G homopolymeric tract) in C. jejuni populations is strongly associated with colonization and enteritis in C57BL/6 IL-10 deficient mice. In the current study, we strengthened the association between locus Cj0170, Cj0045, and mouse colonization. We generated 8 independent strains derived from C. jejuni 11168 strain KanR4 that carried a Cj0170 gene disruption and these were all non motile. Two randomly chosen strains with the Cj0170 gene disruption (DM0170-2 and DM0170-6) were gavaged into mice. DM0170-2 and DM0170-6 failed to colonize mice while the control strain that carried a ''successful''Cj0170 10G allele was motile and did colonize mice. In parallel studies, when we inoculated C. jejuni strain 33292 into mice, the ''unsuccessful''Cj0045 11G allele experienced phase variation to ''successful'' 9G and 10G alleles in 2 independent experiments prior to d4 post inoculation in mice while the ''successful'' 9G allele in the control strain remained stable through d21 post inoculation or shifted to other successful alleles. These data confirm that locus Cj0170 regulates motility in C. jejuni strain KanR4 and is a virulence factor in the mouse model. The data also support a possible role of locus Cj0045 as a virulence factor in strain 33292 in infection of mice.
Disruption of Striated Preferentially Expressed Gene Locus Leads to Dilated Cardiomyopathy in Mice
Liu, Xiaoli,Ramjiganesh, Tripurasundari,Chen, Yen-Hsu,Chung, Su Wol,Hall, Sean R.,Schissel, Scott L.,Padera Jr, Robert F.,Liao, Ronglih,Ackerman, Kate G.,Kajstura, Jan,Leri, Annarosa,Anversa, Piero,Ye Ovid Technologies Wolters Kluwer -American Heart A 2009 CIRCULATION - Vol.119 No.2
<P>BACKGROUND: The striated preferentially expressed gene (Speg) generates 4 different isoforms through alternative promoter use and tissue-specific splicing. Depending on the cell type, Speg isoforms may serve as markers of striated or smooth muscle differentiation. METHODS AND RESULTS: To elucidate function of Speg gene isoforms, we disrupted the Speg gene locus in mice by replacing common exons 8, 9, and 10 with a lacZ gene. beta-Galactosidase activity was detected in cardiomyocytes of the developing heart starting at day 11.5 days post coitum (dpc). beta-Galactosidase activity in other cell types, including vascular smooth muscle cells, did not begin until 18.5 dpc. In the developing heart, protein expression of only Spegalpha and Spegbeta isoforms was present in cardiomyocytes. Homozygous Speg mutant hearts began to enlarge by 16.5 dpc, and by 18.5 dpc, they demonstrated dilation of right and left atria and ventricles. These cardiac abnormalities in the absence of Speg were associated with a cellular hypertrophic response, myofibril degeneration, and a marked decrease in cardiac function. Moreover, Speg mutant mice exhibited significant neonatal mortality, with increased death occurring by 2 days after birth. CONCLUSIONS: These findings demonstrate that mutation of the Speg locus leads to cardiac dysfunction and a phenotype consistent with a dilated cardiomyopathy.</P>