• 제목/요약/키워드: wild type

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Leopard danio 반문의 유전 양상과 생존율에 미치는 영향 (Genetic Analysis of Pigmentation Pattern in the Leopard Danio)

  • 이병문;강거영;송춘복
    • 한국양식학회지
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    • 제11권3호
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    • pp.353-361
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    • 1998
  • Zebra danio와 leopard danio의 부모세대(Po)간의 상반교재를 병행한 교배실험에서 5개체의 small spotted type이 출현한 1개의 실험구를 제외하고는 모두 wild type으로 나타났다. 그리고 F하(1)세대 간의 상반교배로 태어난 F하(2)세대는 wild type과 spotted type의 비율이 모두 3 : 1로 나타났다. F하(1)세대와 Po세대와의 상반교배를 병행한 역교배실험에서는 부모세대를 zebra danio로 사용한 경우, wild type과 spotted type이 178 : 1, 2.7 : 1, 3.9 : 1로 출현하는 실험구를 제외하고는 모두 wild type으로 나타났고, spotted type을 사용한 역교배 및 검정교배실험에서 40개체의 small spotted type으로 출현하는 실험구를 제외하고는 모두 1 : 1의 비율을 나타내었다. Z x F(LZ) 교재실험구 중 두 개의 반복구(각각 2.7 : 1, 3.9 : 1 )에서 wild type과 spotted type의 비율이 3 : 1로 나타났으나, F하(1)세대간의 교배실험과 검정교배실험 결과, zebra danio 암컷이 이형접합자(Pp)였다고 추정되었다. Spotted type과 small spotted type으로 나타나는 예외적인 결과에 대해 아직까지 확실히 규명되지 않았지만, 모든 결과를 종합해 볼 때 반문을 조절하는 유전자의 인자형은 zebra danio가 동형 우성 (PP)이고, leopard danio가 동형 열성(pp)이며, 유전자는 상염색체 상에 존재한다는 것을 알 수 있었다. 수정 후부터 15일째까지의 생존율 비교 실험에서 $Z{\times}Z$ (83.8${\pm}$6.7%)와 $L{\times}L$(80.6${\pm}$4.8%)는 생존율에 있어서 그 차이가 통계적으로 유의하지 않았으며, 이들과 다른 교배군인 $Z{\times}L$(73.2${\pm}$2.0%) 이나 $L{\times}Z$(70.6${\pm}$4.2%) 와는 유의차를 보였다. 따라서 D. frankei로 알려졌던 leopard danio는 zebrafish의 aquarium morph인 반문 돌연변이체이지만 leopard danio 사이에 교배가 이루어질 경우에는 유전적으로 안정되어 있음을 알 수 있는 반면에, wild type zebrafish와는 다소의 유전적인 부조화가 존재한다고 추정된다.

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Comparative lectin binding patterns of Cochlodinium polykrikoides Margalef

  • Rhodes, Lesley L.;Cho, Yong-Chul;Cho, Eun-Seob
    • Journal of the korean society of oceanography
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    • 제35권3호
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    • pp.153-157
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    • 2000
  • Four different FITC-conjugated lectins were used to visually evaluate lectin binding activity by optical staining quality using confocal laser scanning microscopy (CLSM) of Cochzodinium polykrikoides in nature (wild type) and culture (cultured type). Cells from the field and cultures treated with ConA fluoresced only at the outer cell wall, and the abundance and distribution of the fluorescent signal were similar. Treatment with PWM and HPA did not elicit fluorescence at the cell surface, but the wild type exposed to HPA showed greater binding than did the cultured cells, possibly due to greater concentrations of glucosamine. The wild type cells treated with LBL lectin showed a strong green fluorescence on the cell surface, whereas cultured cells did not. Signal intensity and abundance were greater than for any other lectins tested in this study. These results suggest that wild type and cultured type are significantly different based on surface sugar production. In particular, the wild type cells apear richer in galactosamine-like moieties. Neither glucose nor mannose-like moieties were present in either wild types or cultured cells.

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[Retracted] Epinephrine Control of Glycogen Metabolism in Glycogen-associated Protein Phosphatase PP1G/RGLKnockout Mice

  • 김종화;Anna A. DePaoli-Roach
    • BMB Reports
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    • 제35권3호
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    • pp.283-290
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    • 2002
  • The glycogen-associated protein phosphatase (PP1G/$R_{GL}$) may play a central role in the hormonal control of glycogen metabolism in the skeletal muscle. Here, we investigated the in vivo epinephrine effect of glycogen metabolism in the skeletal muscle of the wild-type and $R_{GL}$ knockout mice. The administration of epinephrine increased blood glucose levels from 200±20 to 325±20 mg/dl in both wild-type and knockout mice. Epinephrine decreased the glycogen synthase -/+ G6P ratio from 0.24±0.04 to 0.10±0.02 in the wild-type, and from 0.17±0.02 to 0.06±0.01 in the knockout mice. Conversely, the glycogen phosphorylase activity ratio increased from 0.21±0.04 to 0.65±0.07 and from 0.30±0.04 to 0.81±0.06 in the epinephrine trated wild-type and knockout mice respectively. The glycogen content of the knockout mice was substantially lower (27%) than that of both wild-type mice; and epinephrine decreased glycogen content in the wild-type and knockout mice. Also, in Western blot analysis there was no compensation of the other glycogen targeting components PTG/R5 and R6 in the knockout mice compared with the wild-type. Therefore, $R_{GL}$ is not required for the epinephrine stimulation of glycogen metabolism, and rather another phosphatase and/or regulatory subunit appears to be involved.

Determination and Analysis of Hyper-Variable A Mating Types in Wild Strains of Lentinula edodes in Korea

  • Mi-Jeong Park;Eunjin Kim;Yeun Sug Jeong;Mi-Young Son;Yeongseon Jang;Kang-Hyeon Ka
    • Mycobiology
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    • 제51권1호
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    • pp.26-35
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    • 2023
  • The diversity of A mating type in wild strains of Lentinula edodes was extensively analyzed to characterize and utilize them for developing new cultivars. One hundred twenty-three A mating type alleles, including 67 newly discovered alleles, were identified from 106 wild strains collected for the past four decades in Korea. Based on previous studies and current findings, a total of 130 A mating type alleles have been found, 124 of which were discovered from wild strains, indicating the hyper-variability of A mating type alleles of L. edodes. About half of the A mating type alleles in wild strains were found in more than two strains, whereas the other half of the alleles were found in only one strain. About 90% of A mating type combinations in dikaryotic wild strains showed a single occurrence. Geographically, diverse A mating type alleles were intensively located in the central region of the Korean peninsula, whereas only allele A17 was observed throughout Korea. We also found the conservation of the TCCCAC motif in addition to the previously reported motifs, including ATTGT, ACAAT, and GCGGAG, in the intergenic regions of A mating loci. Sequence comparison among some alleles indicated that accumulated mutation and recombination would contribute to the diversification of A mating type alleles in L. edodes. Our data support the rapid evolution of A mating locus in L. edodes, and would help to understand the characteristics of A mating loci of wild strains in Korea and help to utilize them for developing new cultivars.

Deficiency of iNOS Does Not Prevent Isoproterenol-induced Cardiac Hypertrophy in Mice

  • Cha, Hye-Na;Hong, Geu-Ru;Kim, Yong-Woon;Kim, Jong-Yeon;Dan, Jin-Myoung;Park, So-Young
    • The Korean Journal of Physiology and Pharmacology
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    • 제13권3호
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    • pp.153-159
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    • 2009
  • We investigated whether deficiency of inducible nitric oxide synthase (iNOS) could prevent isoproterenol-induced cardiac hypertrophy in iNOS knockout (KO) mice. Isoproterenol was continuously infused subcutaneously (15 mg/kg/day) using an osmotic minipump. Isoproterenol reduced body weight and fat mass in both iNOS KO and wild-type mice compared with saline-infused wild-type mice. Isoproterenol increased the heart weight in both iNOS KO and wild-type mice but there was no difference between iNOS KO and wild-type mice. Posterior wall thickness of left ventricle showed the same tendency with heart weight. Protein level of iNOS in the left ventricle was increased in isoproterenol-infused wild-type mice. The gene expression of interleukin-6 (IL-6) and transforming growth factor-${\beta}$ (TGF-${\beta}$) in isoproterenol-infused wild-type was measured at 2, 4, 24, and 48-hour and isoproterenol increased both IL-6 (2, 4, 24, and 48-hour) and TGF-${\beta}$ (4 and 24-hour). Isoproterenol infusion for 7 days increased the mRNA level of IL-6 and TGF-${\beta}$ in iNOS KO mice, whereas the gene expression in wild-type mice was not increased. Phosphorylated form of extracellular signal-regulated kinases (pERK) was also increased by isoproterenol at 2 and 4-hour but was not increased at 7 days after infusion in wild-type mice. However, the increased pERK level in iNOS KO mice was maintained even at 7 days after isoproterenol infusion. These results suggest that deficiency of iNOS does not prevent isoproterenol-induced cardiac hypertrophy and may have potentially harmful effects on cardiac hypertrophy.

Habitat preference of wild boar (Sus scrofa) for feeding in cool-temperate forests

  • Kim, Youngjin;Cho, Soyeon;Choung, Yeonsook
    • Journal of Ecology and Environment
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    • 제43권3호
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    • pp.297-304
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    • 2019
  • Background: The growing wild boar population has become a social issue and its feeding characteristics could affect the physical condition and the plant species composition in the South Korean forests. We aimed to reveal the preference of the wild boar on forest type and site condition as feeding grounds in two cool-temperate forested national parks, Odaesan and Seoraksan, in order to provide information to manage the growing population. Results: The 75 plots (53.6%) out of 140 plots were used as feeding grounds by the wild boar, implying a considerably large population. Especially, the observation frequency as feeding ground was the highest in Quercus forests (73.3%), and it was significantly more preferred than deciduous forest type (44.2%) and coniferous forest type (32.4%) (${\chi}^2=17.591$, p < 0.001). Significantly more and deeper pits were found in Quercus forests. Moreover, high elevation and gentle slope ridge were relatively preferred regardless of forest distribution. Conclusions: South Korean forests are growing qualitatively and quantitatively. Particularly, Quercus forest area has increased markedly, while coniferous forest area has decreased. Since the Quercus forest provides rich food sources for the wild boar, the enlargement of this forest type is expected to increase the wild boar population. The forests located at high elevations have high species diversity, and it is expected that these forests will be greatly affected by the increase in the wild boar population as preferred feeding grounds.

Overproduction and Operator DNA-Protein Blotting of R100 Mutant MerR from Shigella flexneri

  • Yoon, Kyung-Pyo
    • Journal of Microbiology and Biotechnology
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    • 제4권4호
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    • pp.250-255
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    • 1994
  • Wild-type and four mutant R100 merR genes were cloned and the proteins overproduced under tac promoter control of pKK223-3. His118Ala, Cys117Ser, Cys126Ser, and wild-type MerR were successfully overproduced although amino-terminal 14 amino acids deletion mutant MerR was not successful. The amount of overproduced wild-type MerR protein as well as other mutant MerR was between 15%-20% of the total protein. The protein was able to be purified up to 95% homogeneity. Specific DNA-protein blotting experiments showed that the 95 bp operator containing DNA fragment could bind to Cys126Ser, His118Ala, and wild- type MerR, but not to Cys117Ser. These results were consistent with the previously reported complementation experiment results that His118Ala, Cys126Ser, and wild-type MerR could repress the mer operon but Cys117Ser could not.

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Site-Directed Mutagenesis of Ile91 of Restriction Endonuclease EcoRV: Dramatic Consequences on the Activity and the Properties of the Enzyme

  • Moon, Byung-Jo;Vipond, I. Barry;Halford, Stephen E.
    • BMB Reports
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    • 제29권1호
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    • pp.17-21
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    • 1996
  • 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.

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Transgenic tobacco plants overexpressing the Nicta; CycD3; 4 gene demonstrate accelerated growth rates

  • Guo, Jia;Wang, Myeong-Hyeon
    • BMB Reports
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    • 제41권7호
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    • pp.542-547
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    • 2008
  • D-type cyclins control the onset of cell division and the response to extracellular signals during the G1 phase. In this study, we transformed a D-type cyclin gene, Nicta;CycD3;4, from Nicotiana tabacum using an Agrobacterium-mediated method. A predicted 1.1 kb cyclin gene was present in all of the transgenic plants, but not in wild-type. Northern analyses showed that the expression level of the Nicta;CycD3;4 gene in all of the transgenic plants was strong when compared to the wild-type plants, suggesting that Nicta;CycD3;4 gene driven by the CaMV 35S promoter was being overexpressed. Our results revealed that transgenic plants overexpressing Nicta;CycD3;4 had an accelerated growth rate when compared to wild-type plants, and that the transgenic plants exhibited a smaller cell size and a decreased cell population in young leaves when compared to wild-type plants.

Microbacterium sp. A-210이 생성하는 Levan fructotransferase의 정제 및 생물학적 특성에 관한 연구 (Purification and Biological Characterization of Wild-type and Mutants of a Levan Fructotransferase from Microbacterium sp. AL-210)

  • 황은영;정미숙;차재호;장세복
    • 생명과학회지
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    • 제19권9호
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    • pp.1218-1225
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    • 2009
  • DFA (Difructose anhydride)는 특유의 구조적인 안정성 때문에 당뇨병 환자를 위한 당원으로써 적합하다는 연구가 보고 되어 있다. DFA에는 4가지 type이 있는데 inulin에 의한 DFA I DFA III DFAV가 있고 levan에 의한 DFA IV가 있는 것으로 알려져 있다. 특히 DFA IV는 당뇨병 환자를 위한 당원 뿐 만 아니라 rat을 이용한 연구에서 칼슘의 흡수를 도와 준다는 보고가 있었다. 이러한 DFAIV를 생성하는 데 쓰이는 Microbacterium sp. AL-210에서 유래한 LFTase (Levan fructotransferase)의 wild-type과 mutants (D63A, D195N, N85S)의 구조적 특성을 밝히기 위해 정제하였다. LFTase의 wild-type과 mutants들을 대량 발현시킨 후 흡착 크로마토그래피, 이온교환 크로마토그래피 그리고 젤 여과 크로마토그래피를 이용하여 고순도로 분리 정제하였으며 이를 SDS-PAGE를 통하여 확인하였다. 분리 정제된 단백질을 JNET 이차 구조 예측 프로그램, solubility 측정, CD (원 편광 이색성 분광편광계), fluorescence spectroscopy (형광분석법), DSC (시차주사열량계)를 이용하여 분석하였다. 또한 다중 정렬과 2차 구조 예측 프로그램을 이용하여 wild-type의 2차 구조를 분석하였다. Solubility 측정에서 가장 적합한 온도는 $55^{\circ}C$, 최상의 pH는 7.5로 나타났다. CD 분석에서 wild-type과 비교한 결과 다른 mutant에 비해 N85S의 $\alpha$-helix가 많이 감소한 것과 $\beta$ strand와 random coil이 증가한 것을 확인하였다. 또한 DSC 분석을 통해 wild-type이 다른 mutants에 비해 안정적인 구조를 지닌 것을 확인하였다. 형광분석에서 N85S가 wild-type과 가장 유사하게 나타났으며 D63A와 D195N은 wild-type에 비해 높은 강도를 나타내었다. 또한 wild-type의 sequence를 Exo-inulinase from Aspegillus awamori, a plant fructan 1-exohydrolase from Cichorium intybus 그리고 invertase from Thermotogo maritime (Tm)의 sequence와 다중 정렬한 결과 Exo-inulinase와 높은 identity를 보였다.