• Title/Summary/Keyword: p104

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Novel p104 protein regulates cell proliferation through PI3K inhibition and p27Kip1 expression

  • Han, Seung-Jin;Lee, Jung-Hyun;Choi, Ki-Young;Hong, Seung-Hwan
    • BMB Reports
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    • v.43 no.3
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    • pp.199-204
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    • 2010
  • The protein p104 was first isolated as a binding partner of the Src homology domain of phospholipase C$\gamma$1, and has been shown to associate with p85$\alpha$, Grb2. The ectopic expression of p104 reduced cellular growth rate, which was also achieved with the overexpression of only the proline-rich region of p104. The proline-rich region of p104 has been found to inhibit the colony formation of platelet-derived growth factor BB-stimulated NIH3T3 cells and MCF7 cancer cells on soft agar. Mutagenesis analysis showed that the second and third proline-rich regions are essential for growth control, as well as for interaction with p85$\alpha$. Overexpression of p104 increased the level of the cyclin-dependent kinase inhibitor, $p27^{Kip1}$, and inhibited the activity of phosphoinositide 3-kinase (PI3K). In summary, p104 interacts with p85$\alpha$ and is involved in the regulation of $p27^{Kip1}$ expression for the reduction of cellular proliferation.

Cloning and Nucleotide Sequence Analysis of the asd Gene from Shigella sonnei KNIH104S (Shigella sonnei KNIH104S로부터 asd 유전자의 클로닝 및 염기서열 분석)

  • 박용춘;신희정;김영창
    • Korean Journal of Microbiology
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    • v.35 no.1
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    • pp.13-17
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    • 1999
  • Shigella sonnez is important causes of human enleric infcctions. S. sonnei KNIH104S was isolated from patient of shigellosis in Korea and previously reported. We cloned 1.7 kb BamHI fragment containing the asd gene encoding an aspartate $\beta$-semialdehyde dehydrogenase from chromosomal DNA of S. sonnei KNIH104S. This recombinant plasmid was named as pSAB17. E. coli $\chi$6097, an a d mutant, cannol grow on the LB medium without DL-$\alpha$, $\varepsilon$-diaminopimclic acid (50 pgiml) but E. coli x 6097(pSAB17) can grow on the same medium. We sequenccd the asd gene ol Shigella for the first time. The asd gcne was composed of 1,104 base pairs with ATG initiation codon and TAA termination codon. Sequence comparison of the asd gene exhibited 99.9% nucleolide sequence hornology with that of E. coli. Also, We constructed the balanced-lethal vector using pBluescrip SK(+) and asd gene of S. sonnei KNIH104S.

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Decolorization of Acid Green 25 by Surface Display of CotA laccase on Bacillus subtilis Spores

  • Park, Jong-Hwa;Kim, Wooil;Lee, Yong-Suk;Kim, June-Hyung
    • Journal of Microbiology and Biotechnology
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    • v.29 no.9
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    • pp.1383-1390
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    • 2019
  • In this study, we expressed cotA laccase from Bacillus subtilis on the surface of B. subtilis spores for efficient decolorization of synthetic dyes. The cotE, cotG, and cotY genes were used as anchoring motifs for efficient spore surface display of cotA laccase. Moreover, a $His_6$ tag was inserted at the C-terminal end of cotA for the immunological detection of the expressed fusion protein. Appropriate expression of the CotE-CotA (74 kDa), CotG-CotA (76 kDa), and CotY-CotA (73 kDa) fusion proteins was confirmed by western blot. We verified the surface expression of each fusion protein on B. subtilis spore by flow cytometry. The decoloration rates of Acid Green 25 (anthraquinone dye) for the recombinant DB104 (pSDJH-EA), DB104 (pSDJH-GA), DB104 (pSDJH-YA), and the control DB104 spores were 48.75%, 16.12%, 21.10%, and 9.96%, respectively. DB104 (pSDJH-EA) showed the highest decolorization of Acid Green 25 and was subsequently tested on other synthetic dyes with different structures. The decolorization rates of the DB104 (pSDJH-EA) spore for Acid Red 18 (azo dye) and indigo carmine (indigo dye) were 18.58% and 43.20%, respectively. The optimum temperature for the decolorization of Acid Green 25 by the DB104 (pSDJH-EA) spore was found to be $50^{\circ}C$. Upon treatment with known laccase inhibitors, including EDTA, SDS, and $NaN_3$, the decolorization rate of Acid Green 25 by the DB104 (pSDJH-EA) spore decreased by 23%, 80%, and 36%, respectively.

The Role of Residues 103, 104, and 278 in the Activity of SMG1 Lipase from Malassezia globosa: A Site-Directed Mutagenesis Study

  • Lan, Dongming;Wang, Qian;Popowicz, Grzegorz Maria;Yang, Bo;Tang, Qingyun;Wang, Yonghua
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1827-1834
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    • 2015
  • The SMG1 lipase from Malassezia globosa is a newly found mono- and diacylglycerol (DAG) lipase that has a unique lid in the loop conformation that differs from the common alpha-helix lid. In the present study, we characterized the contribution of three residues, L103 and F104 in the lid and F278 in the rim of the binding site groove, on the function of SMG1 lipase. Site-directed mutagenesis was conducted at these sites, and each of the mutants was expressed in the yeast Pichia pastoris, purified, and characterized for their activity toward DAG and p-nitrophenol (pNP) ester. Compared with wild-type SMG1, F278A retained approximately 78% of its activity toward DAG, but only 11% activity toward pNP octanoate (pNP-C8). L103G increased its activity on pNP-C8 by approximately 2-fold, whereas F104G showed an approximate 40% decrease in pNP-C8 activity, and they both showed decreased activity on the DAG emulsion. The deletion of 103-104 retained approximately 30% of its activity toward the DAG emulsion, with an almost complete loss of pNP-C8 activity. The deletion of 103-104 showed a weaker penetration ability to a soybean phosphocholine monolayer than wild-type SMG1. Based on the modulation of the specificity and activity observed, a pNP-C8 binding model for the ester (pNP-C8, N102, and F278 form a flexible bridge) and a specific lipid-anchoring mechanism for DAG (L103 and F104 serve as "anchors" to the lipid interface) were proposed.

The Overexpression of Subtilisin Enzyme Using Mutations on Transition State Regulatory Proteins of AprE Promoter and Development of Bacillus subtilis Host System (AprE Promoter전이상태 조절인자 변이주를 이용한 공업적 효소의 과발현과 고초균 숙주계의 개발)

  • 류성호;박승환김병기
    • KSBB Journal
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    • v.11 no.1
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    • pp.8-14
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    • 1996
  • Bacillus subtillis strains with transition state regulator mutations and a spore mutation were developed for the overexpression of apsE and for the enhancement of expression level. Among the many regulator genes, degU and hpr were chosen as a representative positive and negative regulator for the aprE, respectively. Spo II G was used for the construction of asporogeneous strains. All the mutants were constructed from two protease-deleted strain DB104 and the apsE gene was transformed with an integration vector pMK101. DB104(deg$U^h$(32) $his^+$)::pMK101(Cm) and DB104($\Delta$her(Em))::pMKl01(Cm) show 7-fold and about 2-fold increase in aprE expression level, respectively. But the effect of transition state regulator mutation on the aprE expression was diminished when the integrated aprE gene was amplified by the high concentration of chloramphenicol, i. e. 30 $\mu\textrm{g}$/ml. DB104($\Delta$spoIIG(Pm) degUh(32) his+)::pMK101(Cm) and DB104($\Delta$spoIIG(Pm) $\Delta$hpr(Em))::pMK101 double mutant show 10-fold and 3-fold increase in aprE expression level, respectively. The results suggest that sporulation mutation and transition state regulator mutation have independent and additive effect on the aprE expression, and the same gene dosage effect on the transition state regulator mutation was also identified.

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Solubilization of Insoluble Phosphates by Aspergillus sp. PS-104 Isolated from Soil (토양에서 분리한 Aspergillus sp. PS-104 균주에 의한 난용성 인산염 분해)

  • Kang, Sun-Chul;Shin, Seung-Yong
    • Korean Journal of Environmental Agriculture
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    • v.26 no.1
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    • pp.36-41
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    • 2007
  • Phosphate-solubilizing microorganisms were isolated from soil around Kyungnam and Kyungbook regions using potato dextrose agar-calcium phosphate medium. A fungus with the greatest phosphate-solubilizing activity was selected and identified to Aspergillus sp. PS-104, based on the morphological characteristics of conidiophore and conidia; unbranching type of conidiophore, terminally swelling of conidiophore and septate of mycelium, in malt extract agar and potato dextrose agar media. The optimum temperature and initial pH to solubilize rock phosphate in potato dextrose broth-rock phosphate medium were $30^{\circ}C$ and pH 7.0, respectively. In these optimum conditions, phosphate-solubilizing activities of Aspergillus sp. PS-104 against four twos of insoluble phosphate, tricalcium phosphate, aluminium phosphate, hydroxyapatite and rock phosphate, were quantitatively determined. As result, the maximum phosphate-solubilizing activity was obtained with tricalcium-phosphate (1,900 ppm) while minimum activity was obtained with hydroxyapatite (320 ppm). Futhermore, phosphate-solubilizing activity of Aspergillus sp. PS-104 was found higher when treated with nitrates as compared to the ammonium salts as a nitrogen sources.

The Comparison of Psycho-Social Behavior Characteristics between Girls with Precocious Puberty and Normal Girls (성조숙증 여아와 정상발달 여아의 심리사회적 행동특성 비교)

  • Moon, Woo-Jin;Kwon, Ho-Jang;Hwang, Man-Ki
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.2
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    • pp.357-369
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    • 2018
  • This study was conducted to provide data pertaining to precocious puberty by comparing the psycho-social behavior of girls with precocious puberty to those with normal development. This study was conducted from 1 June 2016 to 25 February 2017. The subjects were 104 female patients of A group with precocious puberty visiting H Oriental medicine clinic and S clinic in Seoul, and 104 girls in control group A and 104 girls in control group B attending elementary school in gun. The psycho-social behavioral characteristics of girls with precocious puberty and those with normal development were compared among 312 girls matched for gender and age using a t-test, ${\chi}^2test$, and ANOVA. In addition, the factors influencing precocious puberty were analyzed through multinomial logistic regression. The results revealed that the primary influence factors were frequency of meat intake (p<0.01) and eating-out (p<0.05). These were followed by watching TV (p<0.001), hours of using smart phone (p<0.01) and number of private institutes attended (p<0.05). Additionally, emotional and physical indexes were lower in the precocious puberty group than the control group, indicating that they have more pathology.Finally, girls in the precocious puberty group have lower family and friendship indexes than those in the control group, which means they have more pathology. Overall, the results indicate that extensive research on the causes and frequency of precocious puberty is necessary.

Convergent Study on Microbial Contamination of Toothbrushes According to Intraoral Bacteria Acidogenicity and Toothbrush Care (구강내 세균의 산생성도와 칫솔 관리법에 따른 칫솔 세균 오염에 관한 융합연구)

  • Park, Young-Nam;Ryu, Jae-ki
    • Journal of Convergence for Information Technology
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    • v.10 no.8
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    • pp.223-228
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    • 2020
  • Objective of this study was to compared the amount of bacteria in the toothbrush according to the management method of the toothbrush in use and the acidity of intraoral bacteria. Toothbrushes in use in more than one month of 50 healthy adults were collected and the number of bacteria and pH of the toothbrushes were measured. When the number of brushings per day was 4, the pH 4.97 and the microbial numbers was the lowest at 42.16(104×CFU/ml). When the replacement period of the toothbrush was 1 month, the pH 5.35 and the microbial numbers was the lowest at 19.80(104×CFU/ml). When stored in the bathroom, the pH 4.78, and the microbial numbers was highest at 149.46(104×CFU/ml). As a result, in order to block the germs of the toothbrush, it is necessary to develop a method that can easily disinfect the contaminated toothbrush at home. In addition, I think that it is necessary to educate about the proper brush cleaning and storage method when teaching brushing.