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Shuttle Vectors Derived from pN315 for Study of Essential Genes in <i>Staphylococcus aureus</i>
Matsuo, Miki,Kurokawa, Kenji,Lee, Bok-Luel,Sekimizu, Kazuhisa Pharmaceutical Society of Japan 2010 BIOLOGICAL & PHARMACEUTICAL BULLETIN Vol.33 No.2
<P>Using the <I>par</I> to <I>rep</I> region of the 24653 bp plasmid pN315, which is present in <I>Staphylococcus aureus</I> strain N315, we constructed three vectors that can be shuttled between <I>Escherichia coli</I> and <I>S. aureus</I> and maintained stably in <I>S. aureus</I>. Due to plasmid incompatibility, the resident plasmid in <I>S. aureus</I> cells can be replaced <I>via</I> transformation with an entering plasmid, which carries a different drug resistance gene. To evaluate the applicability of this plasmid-based approach for identifying genes essential for <I>S. aureus</I> cell growth, the chromosomal <I>mraY</I> gene, which is involved in peptidoglycan biosynthesis, was deleted in cells harboring a resident plasmid with an intact <I>mraY</I> gene. The resultant disruptant was then transformed with an empty vector. Cells with a chromosomal <I>mraY</I> deletion but lacking the plasmid supplying <I>mraY</I> could not be recovered, suggesting that <I>mraY</I> is indispensable for staphylococcal cell growth or viability. In contrast, other two genes were shown to be dispensable by this system. Thus, the pN315-based plasmids appear to be useful for studying genes essential for <I>S. aureus</I> cell growth.</P>
Togashi Kenji,Adachi Kazunori,Kurogi Kazuhito,Yasumori Takanori,Watanabe Toshio,Toda Shohei,Matsubara Satoshi,Hirohama Kiyohide,Takahashi Tsutomu,Matsuo Shoichi 아세아·태평양축산학회 2022 Animal Bioscience Vol.35 No.6
Objective: A formula is needed that is practical for current livestock breeding methods and that predicts the approximate rate of inbreeding (ΔF) in populations where selection is performed according to four-path programs (sires to breed sons, sires to breed daughters, dams to breed sons, and dams to breed daughters). The formula widely used to predict inbreeding neglects selection, we need to develop a new formula that can be applied with or without selection. Methods: The core of the prediction is to incorporate the long-tern genetic influence of the selected parents in four-selection paths executed as sires to breed sons, sires to breed daughters, dams to breed sons, and dams to breed daughters. The rate of inbreeding was computed as the magnitude that is proportional to the sum of squared long-term genetic contributions of the parents of four-selection paths to the selected offspring. Results: We developed a formula to predict the rate of inbreeding in populations undergoing four-path selection on genomically enhanced breeding values and with discrete generations. The new formula can be applied with or without selection. Neglecting the effects of selection led to underestimation of the rate of inbreeding by 40% to 45%. Conclusion: The formula we developed here would be highly useful as a practical method for predicting the approximate rate of inbreeding (ΔF) in populations where selection is performed according to four-path programs. Objective: A formula is needed that is practical for current livestock breeding methods and that predicts the approximate rate of inbreeding (∆F) in populations where selection is performed according to four-path programs (sires to breed sons, sires to breed daughters, dams to breed sons, and dams to breed daughters). The formula widely used to predict inbreeding neglects selection, we need to develop a new formula that can be applied with or without selection.Methods: The core of the prediction is to incorporate the long-tern genetic influence of the selected parents in four-selection paths executed as sires to breed sons, sires to breed daughters, dams to breed sons, and dams to breed daughters. The rate of inbreeding was computed as the magnitude that is proportional to the sum of squared long-term genetic contributions of the parents of four-selection paths to the selected offspring.Results: We developed a formula to predict the rate of inbreeding in populations undergoing four-path selection on genomically enhanced breeding values and with discrete generations. The new formula can be applied with or without selection. Neglecting the effects of selection led to underestimation of the rate of inbreeding by 40% to 45%.Conclusion: The formula we developed here would be highly useful as a practical method for predicting the approximate rate of inbreeding (ΔF) in populations where selection is performed according to four-path programs.
Kurokawa, Kenji,Hamamoto, Hiroshi,Matsuo, Miki,Nishida, Satoshi,Yamane, Noriko,Lee, Bok Luel,Murakami, Kazuhisa,Maki, Hideki,Sekimizu, Kazuhisa American Society for Microbiology 2009 Antimicrobial Agents and Chemotherapy Vol.53 No.9
<B>ABSTRACT</B><P>The availability of a silkworm larva infection model to evaluate the therapeutic effectiveness of antibiotics was examined. The 50% effective doses (ED50) of d-cycloserine against the <I>Staphylococcus aureus ddlA</I> mutant-mediated killing of larvae were remarkably lower than those against the parental strain-mediated killing of larvae. Changes in MICs and ED50 of other antibiotics were negligible, suggesting that these alterations are d-cycloserine selective. Therefore, this model is useful for selecting desired compounds based on their therapeutic effectiveness during antibiotic development.</P>
Hirao Akira,Tsunoda Yuji,Matsuo Akira,Sugiyama Kenji,Watanabe Takumi The Polymer Society of Korea 2006 Macromolecular Research Vol.14 No.3
Dendritic hyperbranched poly(methyl methacrylate)s (PMMA)s, whose branched architectures resemble the 'dendron' part(s) of dendrimer, were synthesized by an iterative methodology consisting of two reactions in each iteration process: (a) a coupling reaction of u-functionalized, living, anionic PMMA having two tert-butyldimethylsilyloxymethylphenyl(SMP) groups with benzyl bromide(BnBr)-chain-end-functionalized PMMA, and (b) a transformation reaction of the introduced SMP groups into BnBr functionalities. These two reactions, (a) and (b), were repeated three times to afford a series of dendron-like, hyperbranched (PMMA)s up to third generation. Three dendron-like, hyperbranched (PMMA)s different in branched architecture were also synthesized by the same iterative methodology using a low molecular weight, functionalized 1,1-diphenylalkyl anion prepared from sec-BuLi and 1,1-bis(3-tert-butyldime-thylsilyloxymethylphenyl)ethylene in the reaction step (b) in each iterative process. Furthermore, structurally similar, dendron-like, hyperbranched block copolymers could be successfully synthesized by the iterative methodology using $\alpha$-functionalized, living, anionic poly(2-(perfluorobutyl) ethyl methacrylate) (PRfMA) in addition to $\alpha$-functionalized, living PMMA. Accordingly, the resulting block copolymers were comprised of both PMMA and PRfMA segments with different sequential orders. After the block copolymers were cast into films and annealed, their surface structures were characterized by angle-dependent XPS and contact angle measurements. All three samples showed significant segregation and enrichment of PRfMA segments at the surfaces.
Oku, Yusuke,Kurokawa, Kenji,Matsuo, Miki,Yamada, Sakuo,Lee, Bok-Luel,Sekimizu, Kazuhisa American Society for Microbiology 2009 Journal of Bacteriology Vol.191 No.1
<B>ABSTRACT</B><P>Lipoteichoic acid (LTA) is one of two anionic polymers on the surface of the gram-positive bacterium <I>Staphylococcus aureus</I>. LTA is critical for the bacterium-host cell interaction and has recently been shown to be required for cell growth and division. To determine additional biological roles of LTA, we found it necessary to identify permissive conditions for the growth of an LTA-deficient mutant. We found that an LTA-deficient <I>S. aureus</I> Δ<I>ltaS</I> mutant could grow at 30°C but not at 37°C. Even at the permissive temperature, Δ<I>ltaS</I> mutant cells had aberrant cell division and separation, decreased autolysis, and reduced levels of peptidoglycan hydrolases. Upshift of Δ<I>ltaS</I> mutant cells to a nonpermissive temperature caused an inability to exclude Sytox green dye. A high-osmolarity growth medium remarkably rescued the colony-forming ability of the Δ<I>ltaS</I> mutant at 37°C, indicating that LTA synthesis is required for growth under low-osmolarity conditions. In addition, the Δ<I>ltaS</I> mutation was found to be synthetically lethal with the Δ<I>tagO</I> mutation, which disrupts the synthesis of the other anionic polymer, wall teichoic acid (WTA), at 30°C, suggesting that LTA and WTA compensate for one another in an essential function.</P>
Nakao, Makoto,Hosono, Satoyo,Ito, Hidemi,Oze, Isao,Watanabe, Miki,Mizuno, Nobumasa,Yatabe, Yasushi,Yamao, Kenji,Niimi, Akio,Tajima, Kazuo,Tanaka, Hideo,Matsuo, Keitaro Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.7
Background: Cigarette smoking is a well-established risk factor of pancreatic cancer (PC). Although an association between nicotine dependence phenotype, namely time to first cigarette (TTFC) after waking, and the risk of several smoking-related cancers has been reported, an association between TTFC and PC risk has not been reported. We assessed the impact of smoking behavior, particularly TTFC, on PC risk in a Japanese population. Materials and Methods: We conducted a case-control study using 341 PC and 1,705 non-cancer patients who visited Aichi Cancer Center in Nagoya, Japan. Exposure to risk factors, including smoking behavior, was assessed from the results of a self-administered questionnaire. The impact of smoking on PC risk was assessed with multivariate logistic regression analysis adjusted for potential confounders to estimate odds ratios (ORs) and 95% confidence intervals (CIs). Results: Cigarettes per day (CPD) and/or smoking duration were significantly associated with PC risk, consistent with previous studies. For TTFC and PC risk, we found only a suggestive association: compared with a TTFC of more than 60 minutes, ORs were 1.15 (95%CI, 0.65-2.04) for a TTFC of 30-60 minutes and 1.35 (95%CI, 0.85-2.15) for that of 0-30 minutes (p trend=0.139). After adjustment for CPD or smoking duration, no association was observed between TTFC and PC. Conclusions: In this study, we found no statistically significant association between TTFC and PC risk. Further studies concerning TTFC and PC risk are warranted.