• Title/Summary/Keyword: ermK

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A Study of Electronic Records Folder Management (전자기록철의 구조와 관리방안 - 영국 ERMS 표준을 중심으로 -)

  • Seol, Moon-Won;Cheon, Kwon-Ju
    • Journal of Korean Society of Archives and Records Management
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    • v.5 no.2
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    • pp.49-72
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    • 2005
  • This study aims to analyze the structures of electronic records classification and suggest managing requirements for electronic folder as basic entity for ERM. This present study begins with analyzing the various types of electronic folders based on the Requirements for Electronic Records Management Systems: Functional Requirements of U.K. It designs some examples of classification structures for clarifying the meaning of the electronic folders, components and markers. Finally, it analyses some implications for korean environments including application of electronic folder concept, principles of folder open and closure, and introduction of electronic part entity for efficient folder management.

Identification and Characterization of a Pantothenate Kinase (PanK-sp) from Streptomyces peucetius ATCC 27952

  • Mandakh, Ariungerel;Niraula, Narayan Prasad;Kim, Eung-Pil;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1689-1695
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    • 2010
  • Pantothenate kinase (PanK) catalyzes the first step in the biosynthesis of the essential and ubiquitous cofactor coenzyme A (CoA) in all organisms. Here, we report the identification, cloning, and characterization of panK-sp from Streptomyces peucetius ATCC 27952. The gene encoded a protein of 332 amino acids with a calculated molecular mass of 36.8 kDa and high homology with PanK from S. avermitilis and S. coelicolor A3(2). To elucidate the putative function of PanK-sp, it was cloned into pET32a(+) to construct pPKSP32, and the PanK-sp was then expressed in E. coli BL21(DE3) as a His-tag fusion protein and purified by immobilized metal affinity chromatography. The enzyme assay of PanK-sp was carried out as a coupling assay. The gradual decrease in NADH concentration with time clearly indicated the phosphorylating activity of PanK-sp. Furthermore, the ca. 1.4-fold increase of DXR and the ca. 1.5-fold increase of actinorhodin by in vivo overexpression of panK-sp, constructed in pIBR25 under the control of a strong $ermE^*$ promoter, established its positive role in secondary metabolite production from S. peucetius and S. coelicolor, respectively.

Metabolic Engineering of Rational Screened Saccharopolyspora spinosa for the Enhancement of Spinosyns A and D Production

  • Jha, Amit Kumar;Pokhrel, Anaya Raj;Chaudhary, Amit Kumar;Park, Seong-Whan;Cho, Wan Je;Sohng, Jae Kyung
    • Molecules and Cells
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    • v.37 no.10
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    • pp.727-733
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    • 2014
  • Spinosyns A and D are potent ingredient for insect control with exceptional safety to non-target organisms. It consists of a 21-carbon tetracyclic lactone with forosamine and tri-Omethylated rhamnose which are derived from S-adenosyl-methionine. Although previous studies have revealed the involvement of metK1 (S-adenosylmethionine synthetase), rmbA (glucose-1-phosphate thymidylyltransferase), and rmbB (TDP-D-glucose-4, 6-dehydratase) in the biosynthesis of spinosad, expression of these genes into rational screened Saccharopolyspora spinosa (S. spinosa MUV) has not been elucidated till date. In the present study, S. spinosa MUV was developed to utilize for metabolic engineering. The yield of spinosyns A and D in S. spinosa MUV was $244mgL^{-1}$ and $129mgL^{-1}$, which was 4.88-fold and 4.77-fold higher than that in the wild-type ($50mgL^{-1}$ and $27mgL^{-1}$), respectively. To achieve the better production; positive regulator metK1-sp, rmbA and rmbB genes from Streptomyces peucetius, were expressed and co-expressed in S. spinosa MUV under the control of strong $ermE^*$ promoter, using an integration vector pSET152 and expression vector pIBR25, respectively. Here-with, the genetically engineered strain of S. spinosa MUV, produce spinosyns A and D up to $372/217mgL^{-1}$ that is 7.44/8.03-fold greater than that of wild type. This result demonstrates the use of metabolic engineering on rationally developed high producing natural variants for the production.

A Comparison of Adult and Pediatric Methicillin-Resistant Staphylococcus aureus Isolates Collected from Patients at a University Hospital in Korea

  • Park, Jin-Yeol;Jin, Jong-Sook;Kang, Hee-Young;Jeong, Eun-Hee;Lee, Je-Chul;Lee, Yoo-Chul;Seol, Sung-Yong;Cho, Dong-Taek;Kim, Jung-Min
    • Journal of Microbiology
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    • v.45 no.5
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    • pp.447-452
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    • 2007
  • In this study, we compared the phenotypic and genotypic characteristics of 138 MRSA isolates obtained from adult and pediatric patients (adult, 50; children, 88). The resistance rates against gentamicin, clindamycin, and ciprofloxacin were much higher in the adult MRSA isolates than in the pediatric MRSA isolates. The ermC gene, which is responsible for inducible clindamycin resistance, was detected in 52(59.1%) of the 88 pediatric MRSA isolates but in only 5(10.0%) of the 50 adult MRSA isolates. MRSA isolates of clonal type ST5 with an integration of SCCmec type II/II variants was the most predominant clone among the adult isolates, while clonal type ST72 with an integration of SCCmec IV/IVA was the most predominant clone among the pediatric MRSA isolates. Staphylococcal enterotoxin A and toxic shock syndrome toxin-1 were prevalent among the adult MRSA isolates but not among the pediatric MRSA isolates. The results of this study demonstrated remarkable differences between adult and pediatric MRSA isolates in terms of their antimicrobial susceptibility profiles, SCCmec type, multilocus sequence type, staphylococcal toxin genes, and erythromycin resistance genes.

Heterologous Production of Streptokinase in Secretory Form in Streptomyces lividans and in Nonsecretory Form in Escherichia coli

  • Kim,, Mi-Ran;Choeng, Yong-Hoon;Chi, Won-Jae;Kang, Dae-Kyung;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.132-137
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    • 2010
  • The skc gene encoding streptokinase (SK) with a molecular mass of approximately 47.4 kDa was cloned from Streptococcus equisimilis ATCC 9542 and heterologously overexpressed in Streptomyces lividans TK24 and E. coli using various strong promoters. When the promoter for sprT [Streptomyces griseus trypsin (SGT)] was used in the host S. lividans TK24, a 47.4-kDa protein was detected along with a smaller hydrolyzed protein (44 kDa), suggesting that posttranslational hydrolysis had occurred as has been reported in other expression systems. The casein/plasminogen plate assay revealed that the plasmid construct containing the SGT signal peptide was superior to that containing the SK signal peptide in terms of SK production. Maximal production of SK was calculated to be about 0.25 unit/ml of culture broth, a value that was five times higher than that obtained with other expression systems using ermE and tipA promoters in the same host. When the skc gene was expressed in E. coli BL21(${\Delta}DE3$)pLys under the control of the T7 promoter, a relatively large amount of SK was expressed in soluble form without hydrolysis. SK activity in E. coli/pET28a-$T7_pSK_m$ was more than 2 units/ml of culture broth, even though about half of the expressed protein formed an inactive inclusion body.

Zusammenfassung Die Rechtsstellung des Personals im Luftrecht (민간항공기종사자(民間航空機從事者)의 법적(法的) 지위(地位))

  • Chun, Sam-Hyun
    • The Korean Journal of Air & Space Law and Policy
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    • v.6
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    • pp.239-266
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    • 1994
  • In den 30er Jahren fing man an, sich Gedanken $\ddot{u}ber$ eine internationale Regelung der Rechtsstellung des Luftfahrtpersonals zu machen, um Konflikte zwischen den verschiedenen nationalen Gesetzgebungen hinsichtlich der Handlungen des Luftfahrtpersonals bei der Aus bung seiner Verrichtungen zu verhindern. Aber bisher gibt es keine ausreichende Erfolg. Aus dem Vorhergehenden ergibt sich, $da{\beta}$ eine internationale Regelung der Rechtsstellung des Personals der Zivilluftfahrt erw nscht $w\ddot{a}re$. In der Praxis wird dies jedoch wegen der verschiedenen Arbeitsgesetzgebungen und der anderen nationalen Bestimmungen schwer durchzuf hren sein. Weil aber das Luftfahrtpersonal, insbesondere das Flugpersonal, eine arbeits - und sozialrechtliche Sonderstellung einnimmt, $w\ddot{a}re$ es zu empfehlen, allgemeine internationale Richtlinie zu schaffen, welche in den nationalen $Gesamtarbeitsvertr\ddot{a}gen$ $n\ddot{a}her$ ausgearbeitet werden $k\hat{o}nnten$, sowohl $f\ddot{u}r$ das Kabinenpersonal als $f\ddot{u}r$ die Piloten. Ferner $d\ddot{u}rfte$ es dringend notwendig sein, die Haftpflicht des Kommandanten ausf hrlicher und einheitlicher zu regeln, weil die Intensivierung des Lufttransports und die Anzahl der Flugg ste $st\ddot{a}ndig$ zunehmen. Hinsichtlich der Befugnisse des Kommandanten im Faile von an Bord des Luftfahrzeuges begangenen strafbaren Handlungen gibt das Abkommen von Tokio einige Bestimmungen. Die zivilrechtlichen Aspekte seiner Stellung sind noch nicht festgelegt. Es $k\ddot{o}nnte$ bei internationalen Regelung eine Vorschrift aufgenommen werden, die es $erm\ddot{o}glicht$, einem anderen als dem Kommandanten aile Vollmachten zu erteilen. $Au{\beta}erdem$ $k\ddot{o}nnten$ Regelungen geschaffen werden, welche die Rolle der $\ddot{u}brigen$ Besatzung bei der $Unterst\ddot{u}zung$ des Kommandanten $n\ddot{a}her$ umschreiben. $K\hat{o}nnte$ eine internationale Regelung in diesen Punkten erzielt werden, so $w\ddot{a}re$ sowohl der Einheitlichkeit der Rechtsvorschriften als auch der Rechtssicherheit gedient.

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Effects of Glycerol and Shikimic Acid on Rapamycin Production in Streptomyces rapamycinicus

  • La, Huyen Thi Huong;Nguyen, Thao Kim Nu;Dinh, Hang Thuy;Nguyen, Quyen Minh Huynh;Nguyen, Minh Hong
    • Microbiology and Biotechnology Letters
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    • v.48 no.3
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    • pp.296-302
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    • 2020
  • Rapamycin, derived from Streptomyces rapamycinicus, is an important bioactive compound having a therapeutic value in managing Parkinson's disease, rheumatoid arthritis, cancer, and AIDS. Because of its pharmaceutical activity, studies over the past decade have focused on the biosynthesis of rapamycin to enhance its yield. In this study, the effect of rapG on rapamycin production was investigated. The rapG expression vector was constructed by utilizing the integration vector pSET152 under the control of the erythromycin resistance gene (ermE), a strong constitutive promoter. The rapamycin yield of wild type (WT) and WT/rapG overexpression mutant strains, under fermentation conditions, was analyzed by high-performance liquid chromatography (HPLC). Our results revealed that overexpression of rapG increased rapamycin production by approximately 4.9-fold (211.4 mg/l) in MD1 containing 15 g/l of glycerol, compared to that of the WT strain. It was also found that Illicium verum powder (10 g/l), containing shikimic acid, enhanced rapamycin production in both WT and WT/rapG strains. Moreover, the amount of rapamycin produced by the WT/rapG strain was statistically higher than that produced by the WT strain. In conclusion, the addition 15 g/l glycerol and 15 g/l I. verum powder produced the optimal conditions for rapamycin production by WT and WT/rapG strains.

Radixin Knockdown by RNA Interference Suppresses Human Glioblastoma Cell Growth in Vitro and in Vivo

  • Qin, Jun-Jie;Wang, Jun-Mei;Du, Jiang;Zeng, Chun;Han, Wu;Li, Zhi-Dong;Xie, Jian;Li, Gui-Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9805-9812
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    • 2014
  • Radixin, a member of the ERM (ezrin-radixin-moesin) family, plays important roles in cell motility, invasion and tumor progression. It is expressed in a variety of normal and neoplastic cells, including many types of epithelial and lymphoid examples. However, its function in glioblastomas remains elusive. Thus, in this study, radixin gene expression was first examined in the glioblastoma cells, then suppressed with a lentivirus-mediated short-hairpin RNA (shRNA) method.We found that there were high levels of radixin expression in glioblastoma U251cells. Radixin shRNA caused down-regulation of radixin gene expression and when radixin-silenced cells were implanted into nude mice, tumor growth was significantly inhibited as compared to blank control cells or nonsense shRNA cells. In addition, microvessel density in the tumors was significantly reduced. Thrombospondin-1 (TSP-1) and E-cadherin were up-regulated in radixin- suppressed glioblastoma U251 cells. In contrast, MMP9 was down-regulated. Taken together, our findings suggest that radixin is involved in GBM cell migration and invasion, and implicate TSP-1, E-cadherin and MMP9 as metastasis-inducing factors.

Characterization of Tetracycline-Resistant Genes of Staphylococci isolates from Human and Marine Environment (사람과 해양환경에서 분리된 Staphylococci의 Tetracycline 내성 유전자의 특성 비교)

  • CHO, Ki-Taek;KIM, Young-Chul;KWON, Woo-Ju;JEONG, Hyun-Do
    • Journal of Fisheries and Marine Sciences Education
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    • v.28 no.1
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    • pp.59-68
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    • 2016
  • For comparison of tetracycline-resistant ($Tc^R$) genes, we obtained 21 and 14 $Tc^R$ Staphylococcus spp. from marine environment and human patient, respectively. Although all isolates from human were identified as Staphylococcus aureus, higher proportion of $Tc^R$ isolates (12 out of 14) from human were utilizing tet(M) gene compared to that of $Tc^R$ isolates (6 out of 21) from marine environment. Additionally, collaborated utilization of tet(M) and erm(A) in $Tc^R-Em^R$ S. aureus in human patient, but not in $Tc^R$ Staphylococcus spp. isolates from marine environment was also characterized. Based on the nucleotide sequence of transposon related to $Tc^R$ gene, we confirmed the origin of tet(M) gene in $Tc^R$ Staphylococci isolated from marine environments and human are derived from Tn916/1545-like and Tn5801 transposon, respectively. It is the first report showing the presence of Tn5801 in all $Tc^R$ S. aureus carrying tet(M) in human patient. Alignment of the fully sequenced tet(M) from marine environmental isolates was also agreed with the determined transposons by showing the genomic mosaic structure composed with three genomic parts from Tn916/1545 and unknown transposons. Genetic characteristics of these tet(M) in environmental isolates were similar to each other but different from those in isolates from human showing only tet(M) from Tn916/5801 type. It may imply the presence of less dramatic communication of antibiotic resistant genes between Staphylococci isolated from marine environment and human.

Improvement of Cyclosporin A Hydroxylation in Sebekia benihana by Conjugational Transfer of Streptomyces coelicolor SCO4967, a Secondary Metabolite Regulatory Gene (Sebekia benihana에서 Streptomyces coelicolor SCO4967 유전자 도입을 통한 하이드록실 사이클로스포린 A의 생전환)

  • Kim, Hyun-Bum;Lee, Mi-Jin;Han, Kyu-Boem;Kim, Eung-Soo
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.475-480
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    • 2010
  • Actinomycetes are Gram-positive soil bacteria and one of the most important industrial microorganisms due to superior biosynthetic capabilities of many valuable secondary metabolites as well as production of various valuable bioconversion enzymes. Among them are cytochrome P450 hydroxylase (CYP), which are hemoproteins encoded by a super family of genes, are universally distributed in most of the organisms from all biological kingdoms. Actinomycetes are a rich source of soluble CYP enzymes, which play critical roles in the bioactivation and detoxification of a wide variety of metabolite biosynthesis and xenobiotic transformation. Cyclosporin A (CyA), one of the most commonly-prescribed immunosuppressive drugs, was previously reported to be hydroxylated at the position of 4th N-methyl leucine by a rare actinomycetes called Sebekia benihana, leading to display different biological activity spectrum such as loss of immunosuppressive activities yet retaining hair growth-stimulating side effect. In order to improve this regio-selective CyA hydroxylation in S. benihana, previously-identified several secondary metabolite up-regulatory genes from Streptomyces coelicolor and S. avermitilis were heterologously overexpressed in S. benihana using an $ermE^*$ promoter-containing Streptomyces integrative expression vector. Among tested, SCO4967 encoding a conserved hypothetical protein significantly stimulated region-specific CyA hydroxylation in S. benihana, implying that some common regulatory systems functioning in both biosynthesis and bioconversion of secondary metabolite might be present in different actinomycetes species.