• Title/Summary/Keyword: Gene Identification

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A STUDY OF NI-RESISTANT BACTERIA ISOLATED FROM GINGIVAL CREVICULAR FLUID ON THE PATIENTS WEARING NI-CR ALLOY PROSTHESIS (IN TERMS OF MOLECULAR BIOLOGICAL ASPECTS) (니켈-크롬 합금 보철물 주위 치은 열구내에서 발견된 니켈 내성 균주에 관한 분자생물학적 연구)

  • Chae Young-Ah;Woo Yi-Hyung;Kwon Kung-Rock
    • The Journal of Korean Academy of Prosthodontics
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    • v.41 no.2
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    • pp.207-222
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    • 2003
  • As a material of metal-ceramic prosthesis, nickel as a form of Ni-Cr alloy has been used for many dental prostheses in many cases. However, several problems in use of the alloy have been revealed (ex ; tissue stimulation, skin allergy, hypersensitivity cytotoxicity and carcinogenecity). Little is known about nickel with respect to the relationship between Ni-prosthesis and gaining of Niresistance in oral microorganisms. The present study was undertaken to check whether use of Ni-prosthesis leads to occurrence of Ni-resistant microorganisms. So this study may suggest the possible relationships between the oral microorganisms and nickel-resistance in oral environment. Bacteria were isolated from the gingival crevicular fluid on the patients wearing Ni-Cr prosthesis. The isolated bacteria were tested fir their Ni-resistance in nickel containing media at different concentration from 3mM to 110mM. E. coli HB101 was used as control. The Ni-resistant bacteria were isolated and biochemically identified. The Ni-resistant bacteria were tested several biochemical, molecular-biological tests. Performed tests were : measuring the growth curve, antibiotic test, growth ability test in liquid media, isolation of the chromosome and plasmid, digestion of DNA by restriction enzyme, electrophoresis of chromosome and plasmid DNA, identification of Ni-resistant genes by the DNA hybridization. The results were as follows 1) The bacteria isolated from gingival crevicular fluid on the patients wearing Ni-Cr alloy prosthesis showed nickel-resistance. 2) The isolated microorganisms grew at nickel containing media of high concentrations (60mM-110mM). 3) Based on the biochemical tests, the isolated microorganisms were identified as Enterococcus faecalis(13 cases), Klebsiella pneumoniae(1 case) and Enterobacter gergoviae(1 case). 4) Enterococcus faecalis expressed not only nickel resistance but also the multi-drug resistance to several antibiotics ; chloramphenicol, kanamicin, streptomycin, lincomycin, clindamycin, However, all strain showed the sensitivity against the tetracycline. 5) DNA hybridization result suggests that there is no homology between the previously known gene of nickel resistance in Klebsiella pneumoniae and chromosomal DNA of Enterococcus faecalis.

Relationship Between The Sasang Constitution and Ace Polymophism (사상체질유형과 ACE(angiotensin converting enzyme) 유전자 Type(polymorphism)과의 상관관계)

  • Choi, S.H.;Yim, Y.B.;Rhee, J.W.;Kim, H.Y.;Kang, C.H.
    • Journal of Sasang Constitutional Medicine
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    • v.10 no.2
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    • pp.283-290
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    • 1998
  • Sasang Constitutional Medicine focuses on the different constitutional manifestations of the individual's nature and emotions. The nature and emotions drive the ascending and descending of Qi in the body. And this dynamics of the Qi's ascent and descent shapes the different types of structures, functions and temperaments. Although Sasang Constitutional Medicine has many advantages, its diagnosis of the constitution still depends on the doctors' own idea and has no objective identification. So many doctors in Korea have been trying to solve this problem. Recently, there are several efforts to find out the relationship between genetic information and constitution. By the way, May, 1998 there is a astonishing report about the gene which determines the human performance, that is ACE(angiotensin converting enzyme). And it suggests that the I allele was associated with improved endurance performance. ACE has three genotype including II, ID and DD. "I" means insertion and "D" means deletion. We determined the type of the Sasang constitution with QSCCII questionaire and the one's ACE genotype with PCR of the 127 people and we discovered the relationship between the constitution and the ACE genotype. The result is as follow. Among 39 people who have the II genotype, 7(18%) belong to Taeum(Taiyin), 9(23.1%) belong to Soyang(Shaoyang) and 23(59%) belong to Soeum(Shaoyin). Among 62 people who have the ID genotype, 18(29%) belong to Taeum(Taiyin), 21(33.9%) belong to Soyang(Shaoyang) and 23(37.1%) belong to Soeum(Shaoyin). Among 26 people who have DD genotype, 11(42.3%) belong to Taeum(Taiyin), 4(15.4%) belong to Soyang(Shaoyang) and 11(42.3%) belong to Soeum(Shaoyin). This data indicates that there are implicable relationship between the Sasang constitution and ACE genotype. Especially people who have II genotype have much possibility to be a Soeum(Shaoyin) person (59%) and Soyang(Shaoyang) person have less possibility to have DD genotype (15.4%). With this conclusion, we suggest further study of relationship between the Sasang constitution and ACE genotype and we think that other polymorphism can be a candidate of the partner of Sasang constitution.

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Cloning and Identification of Essential Residues for Thermostable β-glucosidase (BgIB) from Thermotoga maritima (Thermotoga maritima로부터 고온성 β-glucosidase (BgIB)의 클로닝과 필수아미노산 잔기의 확인)

  • Hong, Su-Young;Cho, Kye-Man;Kim, Yong-Hee;Hong, Sun-Joo;Cho, Soo-Jeong;Cho, Yong-Un;Kim, Hoon;Yun, Han-Dae
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1148-1157
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    • 2006
  • A hyperthermophilic bacterium Thernotoga maritima produced thermostable ${\beta}-glucosidase$. The gene encoding ${\beta}-glucosidase$ from T. maritima MSB8 was cloned and expressed in Escherichia coli. The en-zyme (BgIB) hydrolyzed ${\beta}-glucosidase$ linkages between glucose and alkyl, aryl of saccharide groups such as salicin, arbutin, and $_pNPG$. The insert DNA contained ORF with 2,166 bp encodes a 721 amino acids (calculated molecular mass of 80,964 and pl of 4.93). The amino a.id sequence of BglB showed the similarity to family 3 glycosyl hydrolases. The molecular weight of the enzyme was estimated to be approximately 81kDa by MUG-nondenaturing PAGE (4-methylumbelliferyl 13-D-glucoside-nondenaturing polyacrylamide gel electophoresis) and SDS-PACE. The ${\beta}-glucosidase$ exhibited maximal activity at pH 7.0 and $80^{\circ}C$. By exchanging two possible residues (Glu-232 and Asp-242) to Ala by site-directed mutagenesis method, it was found that these were essential for enzymatic activity.

Protease Properties of Protease-Producing Bacteria Isolated from the Digestive Tract of Octopus vulgaris (Octopus vulgaris의 장관으로부터 분리한 단백질 분해효소 생성 균주와 생성된 효소의 특성)

  • Liu, Qing;Ren, Pei;Piao, Meizi;Yang, Ji-Young
    • Journal of Life Science
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    • v.23 no.12
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    • pp.1486-1494
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    • 2013
  • A high protease-producing strain was isolated and identified from the digestive tract of octopus vulgaris by detecting a hydrolysis circle of protease and its activity. The strain was identified by morphology observation, biochemical experiments, and 16S rRNA sequence analysis. The protease obtained from the strain was purified by a three-step process involving ammonium sulfate precipitation, carboxy methyl-cellulose (CM-52) cation-exchange chromatography, and DEAE-Sephadex A50 anion-exchange chromatography. The properties of protease were characterized as well. The strain Bacillus sp. QDV-3, which produced the highest activity of protease, was isolated. On the basis of the phenotypic and biochemical characterization and 16S rRNA gene-sequencing studies, the isolate was identified as follows: domain: Bacteria; phylum: Firmicutes; class: Bacilli; order: Bacillales; family: Bacillaceae; and genus: Bacillus. The isolate was shown to have a 99.2% similarity with Bacillus flexus. A high active protease designated as QDV-E, with a molecular weight of 61.6 kDa, was obtained. The enzyme was found to be active in the pH range of 9.0-9.5 and its optimum temperature was $40^{\circ}C$. The protease activity retained more than 96% at the temperature of $50^{\circ}C$ for 60 min. Phenylmethylsulfonyl fluoride (PMSF) inhibited the enzyme activity, thus confirming that this protease isolated from Bacillus sp. QDV-3 is an alkaline serine protease. Metal ions, $Mn^{2+}$ and $Mg^{2+}$, were determined to enhance the protease activity, whereas $Ba^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ were found to inactivate the enzyme.

Identification and Functional Analysis of the putAP Genes Encoding Vibrio vulnificus Proline Dehydrogenase and Proline Permease

  • Kim, Hye-Jin;Lee, Jeong-Hyun;Rhee, Jee-Eun;Jeong, Hye-Sook;Choi, Hyun-Kyung;Chung, Hee-Jong;Ryu, Sang-Ryeol;Choi, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.318-326
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    • 2002
  • The pathogenic marine bacterium Vibrio vulnificus is the causative agent of food-borne diseases such as life-threatening septicemia. To better understand this organism's strategies to survive osmotic stress, a mutant that was more sensitive to high osmolarity was screened from a library of mutants constructed by a random transposon mutagenesis. By a transposon-tagging method, putAP genes encoding a proline dehydrogenase and a proline permease were identified and cloned from V. vulnificus. The amino acid sequences deduced from nucleotide sequences of putAP from V. vulnificus were 38 to $59\%$ similar to those of PutA and PutP reported from other Enterobacteriaceae. Functions of putAP genes were assessed by the construction of mutants, whose putAP genes were inactivated by allelic exchanges. When proline as the sole carbon or nitrogen source was used, the putA mutant was not able to grow to the substantial level, revealing the proline dehydrogenase is the only enzyme for metabolic conversion of proline into other amino acids. Although the growth rate of the putP mutant on proline as the sole carbon or nitrogen source was significantly reduced, the mutant still grew. This indicated that at least one more proline permease is produced by V. vulnificus. The putP mutant decreased approximately $2-log_10$ CFU/ml after a hyperosmotic challenge, while the parent strain decreased approximately $l-log_10$ CFU/ml. This result suggests that the gene product of putP contributes to the osmotic tolerance of V. vulnificus.

Anti-Obesity Activity of Euptelea Pleiosperma Ethanol Extract (Euptelea pleiosperma 에탄올 추출물의 항비만 활성)

  • Park, Jung Ae;Jin, Kyong-Suk;Kwon, Hyun Ju;Kim, Byung Woo
    • Microbiology and Biotechnology Letters
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    • v.43 no.4
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    • pp.336-342
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    • 2015
  • Previously, Euptelea pleiosperma was identified as one of the useful sources containing anti-oxidative and anti-inflammatory activities for the first time in our research group. In this study, anti-obesity effect of E. pleiosperma ethanol extract (EPEE) was evaluated by using a pancreatic lipase enzyme inhibition assay and a cell culture model system. EPEE suppressed effectively pancreatic lipase enzyme activity dose dependently. Furthermore, EPEE significantly suppressed adipocyte differentiation, lipid accumulation, triglyceride contents, and triggered lipolysis activity on 3T3-L1 preadipocytes in a dose-dependent manner without cytotoxicity. Anti-adipogenic effect of EPEE was modulated by cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins ${\alpha}(C/EBP{\alpha})$, $C/EBP{\beta}$ and peroxisome proliferator-activated receptor ${\gamma}(PPAR{\gamma})$ gene and protein expressions. Taken together, these results provide the important new insight that E. pleiosperma possesses anti-obesity activities such as pancreatic lipase inhibition, anti-adipogenic, and lipolysis effects. It might be utilized as promising sources in the fields of nutraceuticals. The identification of active compounds that confer anti-obesity activity of EPEE might be needed.

Identification of Quantitative Trait Loci for Resistance to Soybean Cyst Nematode Race 14 (콩시스트 선충 race14에 대한 저항성 유전자좌 구명)

  • Choi, In-Soo;Kim, Yong-Chul
    • Journal of Life Science
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    • v.13 no.4
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    • pp.375-382
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    • 2003
  • A direct and precise explanation of soybean resistance to soybean cyst nematode will be possible only when the individual gene(s) involved in the resistance are tagged. This study was conducted, (1) to identify and localize quantitative trait loci for resistance to soybean cyst nematode race 14 on RAPD map, (2) to identify the magnitude and mode of inheritance for each quantitative trait loci, and (3) to identify the best combinations of quantitative trait loci for resistance to soybean cyst nematode race 14. Thirty markers (29 RAPD and 1 RFLP) showed significant association with resistance to soybean cyst nematode race 14. From MAPMAKER/QTL analysis, we identified two regions (linkage group C-7 and linkage group C-9) for resistance to soybean cyst nematode .ace 14. The first quantitative trait loci that was localized at 6.0 cM from $H06^1$ on linkage group C-7 showed a dominant inheritance mode. However, we can not exclude the possibility of additive inheritance mode. The second quantitative trait loci that was localized between $B15^2$ and $E01^1$ on linkage group C-9 also showed a dominant mode of inheritance. One pair of flanking markers ($H06^1$ and $H06^2$) and B15$^2$ were used for multiple regression analysis. Marker combination that included 2 markers, $B15^2$ and $H06^1$, explained the highest total variance (22.9%) for resistance to soybean cyst nematode race 14. Further localization of genes for resistance to soybean cyst nematode race 14 and examination of interaction between quantitative trait loci will accelerate the exploitation of resistance to soybean cyst nematode.

Spinocerebellar ataxia 7 (SCA7) (척수소뇌성 운동실조증 제7형)

  • Seon-Yong, Jeong;Seok-Hun, Jang;Hyon-J., Kim
    • Journal of Genetic Medicine
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    • v.4 no.1
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    • pp.22-37
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    • 2007
  • The autosomal dominant spinocerebellar ataxias (SCAs) are a group of neurodegenerative diseases, clinically and genetically heterogeneous, characterized by degeneration of spinocerebellar pathways with variable involvement of other neural systems. At present, 27 distinct genetic forms of SCAs are known: SCA1-8, SCA10-21, SCA23, SCA25-28, DRPLA (dentatorubral-pallidoluysian atrophy), and 16q-liked ADCA (autosomal dominant cerebellar ataxia). Epidemiological data about the prevalence of SCAs are restricted to a few studies of isolated geographical regions, and most do not reflect the real occurrence of the disease. In general a prevalence of about 0.3-2 cases per 100,000 people is assumed. As SCA are highly heterogeneous, the prevalence of specific subtypes varies between different ethnic and continental populations. Most recent data suggest that SCA3 is the commonest subtype worldwide; SCA1, SCA2, SCA6, SCA7, and SCA8 have a prevalence of over 2%, and the remaining SCAs are thought to be rare (prevalence <1%). In this review, we highlight and discuss the SCA7. The hallmark of SCA7 is the association of hereditary ataxia and visual loss caused by pigmentary macular degeneration. Visual failure is progressive, bilateral and symmetrical, and leads irreversibly to blindness. This association represents a distinct disease entity classified as autosomal dominant cerebellar ataxia (ADCA) type II by Harding. The disease affectsprimarily the cerebellum and the retina by the moderate to severe neuronal loss and gliosis, but also many other central nervous system structures as the disease progresses. SCA7 is caused by expansion of an unstable trinucleotide CAG repeat in the ATXN7 gene encoding a polyglutamine (polyQ) tract in the corresponding protein, ataxin-7. Normal ATXN7 alleles contain 4-35 CAG repeats, whereas pathological alleles contain from 36->450 CAG repeats. Immunoblott analysis demonstrated that ataxin-7 is widely expressed but that expression levels vary among tissues. Instability of expanded repeats is more pronounced in SCA7 than in other SCA subtypes and can cause substantial lowering of age at onset in successive generations termed ‘anticipation’ so that children may become diseased even before their parents develop symptoms. The strong anticipation in SCA7 and the rarity of contractions should have led to its extinction within a few generations. There is no specific drug therapy for this neurodegenerative disorder. Currently, therapy remains purely symptomatic. Cellular models and SCA7 transgenic mice have been generated which constitute valuable resources for studying the disease mechanism. Understanding the pathogenetic mechanisms of neurodegeneration in SCAs should lead to the identification of potential therapeutic targets and ultimately facilitate drug discovery. Here we summarize the clinical, pathological, and genetic aspects of SCA7, and review the current understanding of the pathogenesis of this disorder. Further, we also review the potential therapeutic strategies that are currently being explored in polyglutamine diseases.

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Carbapenemase-Producing Klebsiella oxytoca Detection Using Molecular Methods (분자학적 방법을 이용한 Carbapenemase-Producing Klebsiella oxytoca 검출)

  • Yang, Byoung Seon;Park, Ji Ae
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.4
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    • pp.428-435
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    • 2019
  • The rapid increase and dissemination of carbapene mases, such as Klebsiella pneumoniae carbapenemase (KPC), has become a major problem within the field of healthcare-related infection. There are few antibiotics to treat carbapenem-resistant Enterobacteriaceae (CRE) infections, so the identification of resistant bacterial mechanisms is critical to initiate infection control and conduct epidemiological research. A rapid and effective method for detecting KPC-producing bacteria is needed to avoid therapeutic failures and introduce measures to prevent and control the dissemination of these multi-resistant bacteria. During the study period, 31 isolates (seven isolates of Acinetobacter spp., six isolates of Morganella morganii, five isolates of Pseudomonas aeruginosa, five isolates of Proteus mirabilis, one isolate of Proteus vulgaris, two isolates of Enterobacter cloacae, one isolate of Enterobacter aerogenes, one isolate of Klebsiella pneumoniae, one isolate of Klebsiella oxytoca, one isolate of Serratia marcescens and one isolate of Escherichia coli) were identified by the VITEK. Gram negative rod bacteria were the most frequently isolated from urine (35.5%), blood (19.4%), sputum (16.1%), pus (9.7%), ascitic fluid (9.7%), tracheal aspirates (6.5%) and bile juice (3.2%). Analysis using the PCR method identified the blaKPC gene in the K. oxytoca1 strain, but the blaIMP, blaVIM and blaOXA-48 genes are not amplified. In conclusion, diagnosis using the PCR method can accurately and quickly diagnose KPC, thus establishing quick preventive measures to prevent the spread of KPC in hospitals.

Identification of Korean Mountain Cultivated Ginseng by RAPD (RAPD(Random Amplified Polymorphic DNA)를 이용한 장뇌삼의 지역별 품종 구분)

  • Choi, Ji-Young;Lee, Ju-Hee;Lee, Su-Gwang;Kang, Ho-Duck
    • Journal of agriculture & life science
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    • v.43 no.6
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    • pp.35-43
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    • 2009
  • This study was conducted to examine the genetic variations and intraspecific relationships between 9 individuals of Panax ginseng C.A Meyer by using RAPD (Randomly Amplified Polymorphic DNA) analysis. The 34 primers out of 40 random primers were amplified for all tested plants. The 48 (40%) among 244 bands derived from 34 primers shown polymorphism, and the 72 (64%) rest of bands showed similar forms. By regional groups Sangju and Andong samples located in Kyungsang buk-do showed a high similarity. However, Punggi located in Kyungsang buk-do showed higher similarity with Jinan's of Junla buk-do. In this way, it did not show that Panax ginseng from the same area has similarities. In future study we need to more specific molecular phylogenetic analysis such as AFLP technology and gene sequencing with nuclear chloroplast DNA in all samples.