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Moon, Byung Jo,Vipond, I. Barry,Halford, Stephen E. 생화학분자생물학회 1979 BMB Reports Vol.29 No.1
Ile91 of restriction endonuclease EcoRV, which has not been known to take part directly in catalytic activity, was substituted with Leu by site-directed mutagenesis. The Ile91Leu mutant shows over 1000-fold less activity than the wild type EcoRV under standard reaction condition. The metal ion dependency of the reaction was altered. In contrast to the wild type EcoRV, the mutant prefers Mn^(2+) to Mg^(2+) as the cofactor. In Mn^(2+) buffer the mutant is as active as the wild type enzyme in Mg^(2+) buffer. Like the wild type enzyme, the mutant shows an unspecific binding of DNA in gel shift experiments. In contrast to the wild type enzyme, the mutant did not cleave at noncognate sites of DNA under star condition.
Moon, Byung Jo,Vipond, I. Barry,Halford, Stephen E 생화학분자생물학회 1980 BMB Reports Vol.29 No.2
Site-directed substitutions were made to change the I1e91 of restriction endonuclease EcoRV to either Val, Ala or Gly to identify the role of I1e91 in recognition and catalysis, since substitution of I1e91 with Leu afforded dramatic effects on the activity and properties of restriction endonuclease EcoRV. These changes alter the size of the hydrophobic side chain at position 91 and thus might have revealed the reason for the altered phenotype of Ile91Leu. However, the properties of Ile91Val and Ile91Ala mutants were much like wild type EcoRV, in both activity and metal ion preference. Ile91Gly had very little activity with either Mg^(2+) or Mn^(2+) as cofactors. To try to understand the unusual Mn^(2+) profile of the Ile91Leu mutant, two double mutants, Ile91Leu;Asp90Asn and Ile91Leu;Glu45Met were created. Both double mutants were seriously disabled by the second amino acid change. IIe91Leu;G1u45Met had some residual activity in the Mn^(2+) reaction buffer, whereas the Ile91Leu;Asp90Asn displayed no detectable activity.
Moon, Byung-Jo,Vipond, I. Barry,Halford, Stephen E. Korean Society for Biochemistry and Molecular Biol 1996 Journal of biochemistry and molecular biology Vol.29 No.2
Site-directed substitutions were made to change the Ile91 of restriction endonuclease EcoRV to either Val, Ala or Gly to identify the role of Ile91 in recognition and catalysis, since substitution of Ile91 with Leu afforded dramatic effects on the activity and properties of restriction endonuclease EcoRV. These changes alter the size of the hydrophobic side chain at position 91 and thus might have revealed the reason for the altered phenotype of Ile91Leu. However, the properties of Ile91Val and Ile91Ala mutants were much like wild type EcoRV, in both activity and metal ion preference. Ile91Gly had very little activity with either $Mg^{2+}$ or $Mn^{2+}$ as cofactors. To try to understand the unusual $Mn^{2+}$ profile of the Ile91Leu mutant, two double mutants, Ile91Leu;Asp90Asn and Ile91Leu;Glu45Met were created. Both double mutants were seriously disabled by the second amino acid change. Ile91Leu;Glu45Met had some residual activity in the $Mn^{2+}$ reaction buffer, whereas the Ile91Leu;Asp90Asn displayed no detectable activity.
Moon, Byung-Jo,Vipond, I. Barry,Halford, Stephen E. Korean Society for Biochemistry and Molecular Biol 1996 Journal of biochemistry and molecular biology Vol.29 No.1
Ile91 of restriction endonuclease EcoRV, which has not been known to take part directly in catalytic activity, was substituted with Leu by site-directed mutagenesis. The Ile91Leu mutant shows over 1000-fold less activity than the wild type EcoRV under standard reaction condition. The metal ion dependency of the reaction was altered. In contrast to the wild type EcoRV, the mutant prefers $Mn^{2+}$ to $Mn^{2+}$ as the cofactor. In $Mn^{2+}$ buffer the mutant is as active as the wild type enzyme in $Mn^{2+}$ buffer. Like the wild type enzyme, the mutant shows an unspecific binding of DNA in gel shift experiments. In contrast to the wild type enzyme, the mutant did not cleave at noncognate sites of DNA under star condition.
Three “DS” - Elements for Successful Weight Loss Outcomes: Role of Healthcare Professionals
( Dror Dicker ),( Assim A Alfadda ),( Pernille Auerbach ),( Ian D Caterson ),( Ada Cuevas ),( Jason Halford ),( Masato Iwabu ),( Jae-Heon Kang ),( Rita Nawar ),( Ricardo Arturo Reynoso Mendoza ),( Nic 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1
Aims: In people with obesity (PwO), body weight loss of ≥5% is generally considered to be clinically meaningful. Some PwO do receive treatment and guidance from healthcare professionals (HCPs), but there remains a substantial unmet medical outcomes and weight maintenance need. To identify aspects that might contribute to a successful weight loss outcome (WLO; ≥5% body weight loss maintained for ≥1 year), we investigated the characteristics and experience of PwO with and without successful WLOs using data from the ACTION-IO study (NCT03584191). Methods: An online survey was completed by adults with obesity and HCPs in 11 countries: Australia, Chile, Israel, Italy, Japan, Mexico, Saudi Arabia, South Korea, Spain, UAE and UK. A successful WLO was defined as ≥5% body weight loss in the past 3 years maintained for ≥1 year. Results: A total of 14,502 PwO completed the survey. General characteristics were similar between those who had a successful WLO (n=1,559; 11%) vs those who had not (n=12,943; 89%): 53% vs 52% were male; the mean age was 49 vs 48 years; the mean number of comorbidities was 2.0 vs 1.8. The mean number of serious weight loss attempts was 4 for both groups. However, more PwO who had a successful WLO weighed themselves every day (20%) compared with those who had not had a successful WLO (10%). In terms of interactions with HCPs, more PwO who had a successful WLO had discussed weight (58%) with an HCP within the past 5 years than those who did not have a successful WLO (53%). In addition, more PwO who had a successful WLO compared with those who did not had been diagnosed with obesity (42% vs 35%) and had subsequent direction through the scheduling of a follow-up appointment (25% vs 21%). Conclusions: A 3D approach from HCPs (diagnosis, discussion and direction) appears to be a key element in facilitating a successful WLO. Neither gender, nor age, nor number of weight loss attempts was associated with a successful WLO.