• Title/Summary/Keyword: Mutant

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Associations of the Porcine Melanocortin-4 Receptor (MC4R) Gene with Growth Traits in Duroc Pigs (듀록 품종의 Melanocortin-4 Receptor(MC4R) 유전자와 성장형질과의 연관성 분석)

  • Cho, K.H.;Kim, M.J.;Choi, B.H.;Jeon, G.J.;Ryu, J.W.;Jung, H.J.;Kim, I.C.;Lee, H.K.;Jeon, G.J.
    • Journal of Animal Science and Technology
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    • v.49 no.4
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    • pp.437-442
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    • 2007
  • The melanocortin-4 receptor(MC4R) is virtually expressed in all brain regions and plays an important role in energy homeostasis in mammals. MC4R has been intensively studied as a trait gene controlling economically important traits, such as growth and feed conversion, etc. Six hundreds and sixty Duroc pigs were genotyped for the MC4R locus and analyzed their relationships with breeding values for average daily gain(ADG), backfat thickness(BF), days to 90kg(D90) and feed conversion(FC). The estimated genotype frequencies for the all Duroc pigs were: 30.8%, 45.2%, 24.1% for AA, AB and BB genotypes, respectively. The mutant A allele was significantly associated with ADG, D90 and BF whereas no significant relationship was found with FC. The change of gene frequencies by generation was shown in both selected and culled groups. These results indicate that the MC4R polymorphism could be integrated in the present selection program to realize a marker-assisted selection for the growth traits of the Duroc population.

Dual regulatory effects of PI(4,5)P2 on TREK-2 K+ channel through antagonizing interaction between the alkaline residues (K330 and R355-357) in the cytosolic C-terminal helix

  • Kim, Sung Eun;Kim, Myoung-Hwan;Woo, Joohan;Kim, Sung Joon
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.6
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    • pp.555-561
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    • 2020
  • TWIK-related two-pore domain K+ channel-2 (TREK-2) has voltage-independent activity and shows additional activation by acidic intracellular pH (pHi) via neutralizing the E332 in the cytoplasmic C terminal (Ct). We reported opposite regulations of TREK-2 by phosphatidylinositol 4,5-bisphosphate (PIP2) via the alkaline K330 and triple Arg residues (R355-357); inhibition and activation, respectively. The G334 between them appeared critical because its mutation (G334A) endowed hTREK-2 with tonic activity, similar to the mutation of the inhibitory K330 (K330A). To further elucidate the role of putative bent conformation at G334, we compared the dual mutation forms, K330A/G334A and G334A/R355-7A, showing higher and lower basal activity, respectively. The results suggested that the tonic activity of G334A owes to a dominant influence from R355-7. Since there are additional triple Arg residues (R377-9) distal to R355-7, we also examined the triple mutant (G334A/R355-7A/R377-9A) that showed tonic inhibition same with G334A/R355-7A. Despite the state of tonic inhibition, the activation by acidic pHi was preserved in both G334A/R355-7A and G334A/R355-7A/R377-9A, similar to the R355-7A. Also, the inhibitory effect of ATP could be commonly demonstrated under the activation by acidic pHi in R355-7A, G334A/R355-7A, and G334A/R355-7A/R377-9A. These results suggest that the putative bent conformation at G334 is important to set the tug-of-war between K330 and R355-7 in the PIP2-dependent regulation of TREK-2.

Immobilization of $\beta-galactosidase$ with High Transgalactosylation Activity from Bacillus sp. A4442 Mutant (변이주 Bacillus sp. A4442가 생산하는 갈락토스 전이활성이 높은 $\beta-galactosidase$의 고정화)

  • Kim, Min-Hong;Jung, Jin;In, Man-Jin
    • Applied Biological Chemistry
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    • v.39 no.5
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    • pp.333-337
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    • 1996
  • For continuous production of galactooligosaccharides(GOS), $\beta-galactosidase$ with h1gh transgalactosylation activity from Bacillus sp. A 4442 was Immobilized onto $Diaion^{TM}$ HPA 75(styrene-divinylbenzene resin). The parameters influencing enzyme immobilization were scrutinized in order to maximize immobilization yield while minimizing enzyme inactivation. The optimum conditions turned out to be: Tris buffer concentration 30 mM, pH 8.0, contact time at room temperature 3 hr, and enzyme loading 25 mg protein/g resin. Both the thermal stability and the operational stability of immobilized enzyme were markedly enchanced by the treatment with 0.5% glutaraldehyde as a cross-linker. Under the experimental conditions established, the yield of ${\beta}-galactosidase$ immobilization was 40% or more and the activity of the immobilized enzyme ca. 200 U/g resin. When a packed-bed reactor was employed to continuously convert lactose to GOS, the specific production, which refers to as the amount of commercially valuable GOS produced by a unit amount of immobilized ${\beta}-galactosidase$, was found to be ca. 300 g GOS/g carrier.

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Production of ${\beta}-Galactosidase$ with High Transgalactosylation Activity by Bacillus sp. A4442 Mutant (변이주 Bacillus sp. A4442에 의한 갈락토스 전이활성이 높은 ${\beta}-Galactosidase$의 생산)

  • Choi, Kyung-Ho;Yang, Sung-Joon;Kim, Min-Hong;Han, Keum-Soo;Yang, Ji-Won;Jung, Jin;In, Man-Jin
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.507-511
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    • 1995
  • In an attempt to improve the productivity of ${\beta}-galactosidase$ from Bacillus sp. A1, which was isolated from soil and has remarkably higher transgalactosylation activity than lactose hydrolysis activity, a chemical mutation procedure using N-methyl-N'-nitro-N-nitrosoguanidine followed by selection was conducted. The final selection, designated as Bacillus sp. A4442, turned out to show a substantially increased enzyme productivity. Catabolite repression by glucose and lactose requirement as an inducer for the enzyme biosynthesis, which were shown in the parent strain, was markedly diminished; instead it was found out that galactose acts as another inducer. Because pH of medium, one of the most important factors for cell growth as well as enzyme production, is closely related with the sugar concentration during culture, it was kept in the optimum range of $6.5{\sim}7.5$; for this the initial glucose concentration was adjusted to be 0.5% which was thereafter maintained by the controlled pumping-in of lactose using the pH-stat technique. By doing so, we were able to increase the productivity of ${\beta}-galactosidase$ with high transgalactosylation activity up to $44\;unit/m{\ell}-broth$.

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Gene Structure and Function of fkhE, a Forkhead Gene in a Filamentous Fungus Aspergillus nidulans (Aspergillus nidulans forkhead 유전자 fkhE의 구조와 기능 분석)

  • Park, Mi-Hye;Kim, Hyoun-Young;Kim, Jong-Hwa;Han, Kap-Hoon
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.160-166
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    • 2010
  • A homothallic filamentous fungus Aspergillus nidulans has been used as the a model organism for studying growth and development for eukaryotic system. Various studies about specific transcription factors have been performed for elucidating the molecular mechanisms of growth, asexual and sexual developmental processes. Among them, the fkhE gene (AN2025.3) is located in chromosome VII and contains an ORF encoding 718 amino acid polypeptide intervening with two short introns. The cDNA sequencing revealed that at least four types of alternative splicing events were occurred when the fkhE gene was transcribed. The putative FkhE polypeptide contains a conserved forkhead domain and a bipartite nuclear localization signal at it's N-terminus and C-terminus, respectively. Deletion of fkhE resulted in impaired conidiophore formation in a solid medium. However, the sexual developmental process or cleistothecia formation was normal. Furthermore, fkhE deletion mutant produced conidiophores and conidia under the submerged culture, indicating that the fkhE gene is involved in asexual developmental process similar to the fkhF gene.

The Developmental Characterization of the Sericin jam(Bombyx mori), a Mutant in the Synthesis and Secretion of Fibroin (세리신잠(Nd-s. $Nd^H$의 발육 특성)

  • 김수연;손해룡
    • Journal of Sericultural and Entomological Science
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    • v.45 no.1
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    • pp.6-9
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    • 2003
  • This experiment was done to know developmental characteristics of Sericin jam. Incubation periods were 10 day 2 hr, and 10 day 1 hr. 11 day 1 hr for Nd-s jam, N $d^{H}$ jam, and Baegok jam, respectively. Hatching rates were 83.9, 83.3 and 96.0% for Nd-s jam, N $d^{H}$ jam, and Baegok jam. Larval periods were, 20 days 1 hr for Nd-s jam, 20 days 5 hrs for N $d^{H}$ jam, and 22 days 12 hrs for Baegok jam. Death rate of larvae was highest in N $d^{H}$ iam, followed by Nd-s jam and Baegok jam. Pupation rate was highest in Baegok Jam followed by Nd-s jam and that of N $d^{H}$ jam was the lowest among the three. Cocoon weight was 1.39, 1.08, and 2.01 g for Nd-s jam, N $d^{H}$ jam, and Baegok jam, respectively. Shell weight were 13, 3, and 48 cg for Nd-s jam, N $d^{H}$ jam, and Baegok jam. Cocoon shell ratios were 9.0% for Nd-s jam, 2.8% for N $d^{H}$ jam and 23.9% for Baegok jam. Cocoon sizes were 30.6${\times}$15.8 mm for Nd-s jam, 24.7${\times}$14.9 mm for N $d^{H}$ jam and 35.8 ${\times}$ 20.5 mm(1${\times}$w) for Baegok jam.w) for Baegok jam.

Effect of Silkworm Hemolymph Collected Large-scale Bleeding Method in Insect Cell Culture (절피원심법으로 대규모 채취한 누에체액의 곤충세포 증식효과)

  • Choi, Ji-Young;Kim, Jong-Gill;Choi, Young-Cheol;Kim, Sam-Eun;Han, Myung-Sae
    • Journal of Sericultural and Entomological Science
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    • v.49 no.1
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    • pp.28-32
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    • 2007
  • This study was carried out to investigate the utilization of hemolymph of silkworm, Bombyx mori, as a substitute for fetal bovine serum(FBS) in the insect cell culture. Hemolymph is collected on a small scale by clipping the abdominal leg; however, this method is not appropriate for large scale collection. The hemolymph was collected from 5 th instar larva by centrifugation after cutting of the abdominal legs was more appropriate procedure for large scale collection. The cell growth in the medium supplemented with hemolymph(Baekokjam) collected in large scale was almost same as that in the medium hemolymph supplemented with hemolymph collected in small scale. However, the mutant($wE^b$) hemolymph collected in large scale was still less effective in the cell growth, as compared to the Baekokjam hemolymph collected in large scale. The optimum centrifugation condition for large-scale bleeding was 500 rpm and 15 min.

Structure-Function Analysis of DNA Binding Domain of the Yeast ABF1 Protein (효모 ABF1 단백질의 DNA Binding 부위에 대한 구조 기능 연구)

  • Cho, Gi-Nam;Lee, Sang-Kyung;Kim, Hong-Tae;Kim, Ji-Young;Rho, Hyune-Mo;Jung, Gu-Hung
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.102-108
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    • 1994
  • Autonomously replicating sequence Binding Factor 1(ABF1) is a DNA-binding protein that specifically recognizes the $RTCRYN_5ACG$ at many sites in the yeast genome including the promoter element, mating-type silencer and ARS. To express the intact full-length ABF1 gene in E. coli, the ABF1 gene has been cloned into pMAL-c2 and His-61, Leu-353 and Leu-360 were substituted with other amino acid. ABF1 fusion proteins of wild type ABF1 and H61A, L353R and L360R nutants were purified by amylose resin affinity chromatography. Fusion protein of MBP and ABF1 was digested by Factor Xa and Characterized by gel retardation assay and complementation test. As aresult, we suggested that other DNA binding motif except atypical inc-finger motif is in the middle region of ABF1.

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Identification and Characterization of the Aquaporin Gene aqpA in a Filamentous Fungus Aspergillus nidulans (사상성 진균 Aspergillus nidulans에서 아쿠아포린 유전자 aqpA의 분리 및 분석)

  • Oh, Dong-Soon;Lu, Han-Yan;Han, Kap-Hoon
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.295-301
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    • 2011
  • Aquaporin is a water channel protein, which is classified as Major Intrinsic Protein (MIP), found in almost all organisms from bacteria to human. To date, more than 200 members of this family were identified. There are two major categories of MIP channels, orthodox aquaporins and aquaglyceroporins, which facilitate the diffusion across biological membranes of water or glycerol and other uncharged compounds, respectively. The full genome sequencing of various fungal species revealed 3 to 5 aquaporins in their genome. Although some functions of aquaporins found in yeast were characterized, however, no functional characteristics were studied so far in filamentous fungi, including Aspergillus sp. In this study, one orthodox aquaporin homolog gene, aqpA, and four aquaglyceroporin homologs, aqpB-E, in a model filamentous fungus Aspergillus nidulans were identified and the function of the aqpA gene was characterized. Knock-out of the aqpA gene didn't show any obvious phenotypic change under the osmotic stress, indicating that the function of the gene does not involved in the osmotic stress response or the function could be redundant. However, the mutant showed antifungal susceptibility resistance phenotype, suggesting that the function of the aqpA gene could be involved in sensing the antifungal substances rather than the osmotic stress response.

Studies of cold resistant glycine betaine effect on cold sensitive Bacillus subtilis mutant strains (저온 민감성 바실러스 서브틸리스 돌연변이 균주에서 glycine betaine의 저온 내성에 미치는 영향에 대한 연구)

  • Kim, Do Hyung;Lee, Sang Soo
    • Korean Journal of Microbiology
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    • v.54 no.3
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    • pp.200-207
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    • 2018
  • At high salt concentration, glycine betaine is transported into Bacillus subtilis and growing rate of the cell is not suppressed. Also according to recent studies, cell growth is maintained normal growth rate at low temperature. Low temperature results in a stress response of Bacillus subtilis that is characterized by strong repression of major metabolic activities such as translation machinery and membrane transport. In this regards, genes showing cold sensitive phenotype are cold-induced DEAD box RNA helicases (ydbR, yqfR) and fatty acid desaturases (bkdR, des). Therefore to understand the effect of glycine betaine on cold growth of Bacillus subtilis, we investigated the effect of glycine betaine on growth rate of these deletion mutants showing cold sensitive phenotype. Glycine betaine strongly stimulated growth of wild type Bacillus subtilis JH642 and deletion mutants of ydbR and yqfR at $20^{\circ}C$ (190~686 min $T_d$ difference). On the other hands, glycine betaine does not show growth promoting effects on deletion mutants of bkdR, and des at cold conditions. Same cold protectant growth results were shown with the precursor choline instead of glycine betaine. We investigated the effects of detergents on the cell membrane in bkdR and des deficient strains associated with cell membrane. It was identified that bkdR deficient strain shows retarded growth with detergent such as Triton X-100 or N-lauryl sarcosine compared with wild type cell. Thus, it is possible that deletion mutation of bkdR modifies membrane structure and effects on transport of glycine betaine.