• Title/Summary/Keyword: Substitution site

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Site-Directed Mutagenesis Studies with Restriction Endonuclease EcoRV to Identify the Role of Ile91 in Recognition and Catalysis

  • Moon, Byung-Jo;Vipond, I. Barry;Halford, Stephen E.
    • BMB Reports
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    • v.29 no.2
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    • pp.99-104
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    • 1996
  • 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.

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Mitochondrial DNA polymorphism in Fourteen Geographical Strains of Drosophila melanogoater (세계 14지역 계통에 대한 초파리 미토콘드리아 DNA의 다형현상)

  • 김봉기
    • The Korean Journal of Zoology
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    • v.31 no.3
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    • pp.218-224
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    • 1988
  • Restriction endonucleases were used to search for intraspecific variation at 32 cleavage sites in mitochondrial DNA(mtDNA) purified from fourteen strains of Drosophila melanogaster helonging to different localities of the world. mtDNA of D. melanogaster was displayed site variation(Hpall, Haelll and Seal endonucleases) and length variation(maxirnum 550bp). Six genotypes, Ml, M2, M3, M4, M6 and M7, could be distinguished based on ihe site types witti a low average of intraspecific substitution rate (1.88%),but M5 type of Ogasawara strain in Japan was not detected in this study. A possible explanation for the low divergence was that mtDNA variation of fourteen strains in D. melanogaster could not he accumulated sufficiently owing to recent divergence of few individuals, and that sequence divergence was prevented by frequent migration in spite of the geographical isolation.

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A Series of Quinoline-2-carboxylic Acid Derivatives: New Potent Glycine Site NMDA Receptor Antagonists

  • 김란희;최진일;최승원;이광숙;정영식;박우규;성철민;박노상
    • Bulletin of the Korean Chemical Society
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    • v.18 no.9
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    • pp.939-945
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    • 1997
  • Several types of 4-substituted-quinoline-2-carboxylic acid derivatives possessing different substituents at C4-position such as sulfonyl, phosphonyl, carbonyl groups, or a flexible alkyl chain have been synthesized and evaluated for their in vitro antagonistic activity at the glycine site on the N-methyl-D-aspartate (NMDA) receptor. Of them, 5,7-dichloro-4-(tolylsulfonylamino)-quinoline-2-carboxylic acid 9 was found to have the best in vitro binding affinity with IC50 of 0.57 μM. On the other hand, in compounds 21 and 22 the introduction of flexible alkyl chains on C4 of the quinoline mother nuclei caused a significant decrease of the in vitro binding affinity. In addition, replacement of polar carboxylic acid group on C2 by neutral bioisosteres in compounds 23a-d also seems to be disadvantageous to in vitro activity. In the structure-activity relationship (SAR) study of the 4-substituted quinoline-2-carboxylic acid acid derivatives, it was realized that the substitution pattern on C4 significantly influences on the binding affinity for the glycine site of NMDA receptor and the binding affinity might be increased by the introduction of a suitable electron rich substituent at C4 which has the ability of H-bonding donor.

Genetic Analysis of Ancient Human Bones Excavated in Sacheon Nuk-do and Gyeongsan Yimdang-dong, Korea (경산 임당동 및 사천 늑도 출토 인골의 유전자 분석)

  • Seo, Min-Seok;Lee, Kyu-Shik
    • 보존과학연구
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    • s.25
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    • pp.47-74
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    • 2004
  • We investigated the nucleotide substitution and insertion polymorphism of the hypervariable region Ⅰ and Ⅱ in mt DNA by sequencing ancient DNA from 51 ancient bones and teeth excavated at Nuk-do and Yimdang-dong in Korea. It revealed 35 sequence types from the ancient Korean. Of these, different sequences were 34 sequences. There were 19 and 38 base substitutions in HVI and HVⅡ, respectively. Some substitutions were characteristic of East Asian populations as compared with data reported on Caucacianpopulations,16051, 16150, 16172, 16223 in region I and 73, 263 in region II were noted as polymorphic sites, respectively. These were distributed evenly along the control region, though the frequency of each site was variable. Nucleotide substitution rather than insertion and deletion was the prevalent pattern of variation. Insertion of cytosine between312 and 315 in region HVⅡ were detected up to 98% in 51 ancient bone samples. This sequence data represents a phylogenetic tree using NTI DNA Suite computer program. The phylogenetic tree showed that mt DNA sequences of Nuk-do bones were relative to west Siberian and Indonesian. The usefulness of mt DNA sequencing in ancient Korean population excavated atarchaeological sites is based on biological and historical evidence for origin and migration of ancient Korean.

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Differential Evolution between Monotocous and Polytocous Species

  • Ahn, Hyeonju;Kim, Kyu-Won;Kim, Hyeon Jeong;Cho, Seoae;Kim, Heebal
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.4
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    • pp.464-470
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    • 2014
  • One of the most important traits for both animal science and livestock production is the number of offspring for a species. This study was performed to identify differentially evolved genes and their distinct functions that influence the number of offspring at birth by comparative analysis of eight monotocous mammals and seven polytocous mammals in a number of scopes: specific amino acid substitution with site-wise adaptive evolution, gene expansion and specific orthologous group. The mutually exclusive amino acid substitution among the 16 mammalian species identified five candidate genes. These genes were both directly and indirectly related to ovulation. Furthermore, in monotocous mammals, the EPH gene family was found to have undergone expansion. Previously, the EPHA4 gene was found to positively affect litter size in pigs and supports the possibility of the EPH gene playing a role in determining the number of offspring per birth. The identified genes in this study offer a basis from which the differences between monotocous and polytocous species can be studied. Furthermore, these genes may harbor some clues to the underlying mechanism, which determines litter size and may prove useful for livestock breeding strategies.

Synthesis and Thermoelectric Properties of the B-Site Substituted SrTiO3 with Vanadium

  • Khan, Tamal Tahsin;Mahmud, Iqbal;Ur, Soon-Chul
    • Korean Journal of Materials Research
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    • v.27 no.8
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    • pp.416-421
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    • 2017
  • V-substituted $SrTiO_3$ thermoelectric oxide materials were fabricated by the conventional solid state reaction method. From X-ray diffraction pattern analysis, it can be clearly seen that almost every vanadium atom incorporated into the $SrTiO_3$ provided charge carriers. The electrical conductivity ${\sigma}$, Seebeck coefficient S, and thermal conductivity k were investigated in a high temperature regime above 1000 K. The addition of vanadium significantly reduced the thermal conductivity and enhanced the Seebeck coefficient, as well as the electrical conductivity, thus enhancing the ZT value. A maximum ZT value of 0.084 at 673 K was observed for the sample with 1.0 mole% of vanadium substitution. In this study, the reason for the enhanced thermoelectric properties via vanadium addition was also investigated.

Reuse of Treated Sewage Water from Absorbent Biofilter System as Agricultural Water Resources (농업수자원으로서의 흡수성 Biofilter 처리수 재이용)

  • 권순국;김현욱;권용웅;조영현;박상원;임경래
    • Magazine of the Korean Society of Agricultural Engineers
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    • v.45 no.5
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    • pp.151-159
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    • 2003
  • Absorbent Biofilter Systems (ABS), composed of an anaerobic septic tank, a pump chamber and an absorbent biofilter tank, have been found to economically provide rural on-site wastewater treatment. This study was conducted to assess the potential of ABS effluent as an alternative water resource for agricultural and environmental use, with respect to the removal of pathogenic microorganism and their fertilization effect. A pilot scale ABS was used to compare its removal efficiency of pathogens from effluent water. Overall, more than 95 percent of Salmonella and E. coli were removed. This result demonstrates that a significant reduction in the pathogenic microorganism of effluents can occur in ABS, which implies the feasibility for the use of ABS effluent in agriculture and environment, with the provision of a further simple disinfection step, in order to satisfy the WHO guidelines for the microbiological quality in agriculture. In addition, because of the abundant nutritional content of ABS effluent, the substitution effect of fertilizer (N, P and K) in paddy irrigation, i.e. 2/3 for nitrogen, l/3 for phosphorus and 1/5 for potassium would be expected. Based on the experimental data, the ABS effluent could be used as a new alternative water resource for paddy irrigation, as well as for environmental purposes, such as supplying water to ecological parks in rural villages.

Theoretical Studies on Electrophilic Substitution of Five-membered Heteroaromatic Compounds with Isopropyl Cation (5각-이종원자 방향족 고리화합물과 이소프로필 양이온간의 친전자 치환반응에 관한 이론적 연구 (1))

  • Lee, Ik Choon;Kim, Chang Gon;Lee, Bon Su
    • Journal of the Korean Chemical Society
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    • v.34 no.3
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    • pp.248-254
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    • 1990
  • Theoretical studies on the electrophilic substitution reactions of five-membered heteroaromatic compounds, furan, pyrrole and thiophene, with isopropyl cation were carried out using the MNDO method. The results indicated that site selectivities of ${\alpha},{\beta}$ and hetero-atoms are not controlled by electrostatic interactions but are determined mainly by charge trasfer stabilization between the HOMO of heteroaromatics and the LUMO of the electrophile. The reactivity order for ${\alpha}\;and\;{\beta}$ positions was pyrrole > furan > thiophene, in agreement with the solution-phase as well as the gas-phase experimental results.

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Oligomerization of the substitution mutants of autographa californica nuclear polyhedrosis Virus (AcNPV) gp64 glycoprotein

  • Kim, Ki-Nam;Poo, Ha-Ryoung;Yang, Jai-Myung
    • Journal of Microbiology
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    • v.35 no.1
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    • pp.72-77
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    • 1997
  • The baculovirus gp64 glycoprotein is a major component of the envelope protein of budded virus (BV). It has been shown that the gp64 glycoprotein plays an essential role in the infection process, especialy fusion between virus envelope and cellular endosomic membrane. Recently we reported optimal conditions required for gp64-mediated membrane fusion in pGP64 DNA transfected Spodoptera frugiperda (Sf9) cells (H. J. Kim and J. M. Yang, Jour, Microbiology, 34.7-14). In order to investigate the role of hydrophobicity within the fusion domain of the gp64 glycoprotein for membrane fusion, 13 mutants which have substitution mutation within hydrophobic region I were constructed by PCR-derived site-derected mutagenesis. Each mutated gp64 glycoproteins was transiently expressed by transfecting plasmid DNA into Spodoptera frugiperda (Sf9) cells. Oligomerization of the transisently expressed gp64 glycoproteins was a nalysed by running them on SDS-polyacrylamide gel electrophoresis under non-reducing condition followed by immunoblotting. All of the mutant gp64 glycoproteins expect cysteine-228 were able to form trimers. These results suggest that hydrophobic region I of the gp64 may not be responsible for the oligomerization of the gp64 glycoprotein.

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The Characteristics of I269S and I224S Double Mutant Horse Liver Alcohol Dehydrogenase (I269S와 I224S 이중변이 알코올 탈수소효소의 특성)

  • Ryu, Ji-Won;Lee, Kang-Man
    • YAKHAK HOEJI
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    • v.41 no.6
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    • pp.756-764
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    • 1997
  • Ile-224 in I269S mutant horse liver alcohol dehydrogenase isoenzyme S (HLADH-S) was mutated to serine by site-directed mutagenesis in order to study the role of the residue in c oenzyme binding to the enzyme. The specific activity of the I269S and I224S mutant enzyme to ethanol was increased 6-fold and all Michaelis constants($K_a,\;K_b,\;K_p,\;and\;K_q$,/TEX>) were larger than those for the wild-type and I269S enzyme. The substitution decreased the afffinity to coenzymes and increased the specific activity of the enzyme. The mutant enzyme showed the highest catalytic efficiency for octanol among the primary alcohols. But it didn`t have activities on retinoids and 5${\beta}$-cholanic acid-3-one. From these results, it was confirmed that the hydrophobic interaction of Ile-224 residue with coenzyme was related to coenzyme affinity in ADH reaction. The substitution also affected the substrate affinities to the enzyme.

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