• Title/Summary/Keyword: 제한효소분석

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Arrest of Cell Growth by Inhibition of Endogenous Reverse Transcription Activity in Cancer and Somatic Cell Lines (사람의 암세포주 및 정상세포주에서 역전사 효소의 억제에 의한 세포 성장의 제한)

  • Mi-Jeong Kim;Sung-Ho Lee;Jong-Kuen Park;Byeong-Gyun Jeon
    • Journal of Life Science
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    • v.34 no.6
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    • pp.365-376
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    • 2024
  • The present study assessed the cytotoxic effects on cell growth and senescence in human cancer (A-549, AGS, HCT-116, MDA-MB-231, and U 87-MG) and normal (MRC-5 and mesenchymal stem cells) cell lines treated with efavirenz (EFA), an inhibitor of non-nucleoside reverse transcriptase (RTase). Following EFA treatment, the half-maximal inhibitory concentration (IC50) values were approximately 15 µM, and the IC50 value was significantly (p<0.05) lower in the cancer cell lines, compared to normal cell lines. After determining the IC50 values against EFA, each cell line was treated with 15 µM EFA for up to one week. Significant (p<0.05) decreases in endogenous RTase and telomerase activity were observed in the cancer cell lines. RTase and telomerase activity were absent or detected at very low levels in both EFA-untreated and treated MRC-5 and MSC normal cells. The cell doubling time (CDT) was also significantly (p<0.05) prolonged by the decreased cell growth rate in the EFA-treated cancer cell lines compared to the untreated cell lines. Furthermore, EFA-treated cancer cells displayed a high number of cells with a high intensity of senescence-associated ß-galactosidase activity (SA-ß-gal activity), compared to the untreated cells. The present study showed that inhibition of RTase activity induces cellular senescence and arrests cell growth in human cancer cell lines; however, normal cell lines showed greater tolerance against EFA. RTase treatment could offer optional chemotherapy for cancer treatment in human cancer cell lines with high RTase activity.

PCR-RFLP Analysis of Ribosomal DNA Intergenic Spacer Region in Fusarium section Liseola. (Fusarium section Liseola 균주들에서 rDNA Intergenic Spacer 부위의 PCR-RFLP 분석)

  • 이경은;최영길;민병례
    • Korean Journal of Microbiology
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    • v.38 no.1
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    • pp.7-12
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    • 2002
  • The intergenic spacer (IGS) region of the ribosomal DNA of species in Fusarium section Liseola was analyzed by amplification and subsequent digestion with several restriction enzymes. The length of the amplified IGS region was about 2.6 Kb in all strains except F.moniliforme 12 Which was about 2.9 Kb. The enzymes, EcoRI, HincII, SalI, HindIII, PstI and SmaI, digested the IGS region and nine haplotypes were identified among 11 strains. In the dendrogram based on PCR-RFLP of IGS region combined the results of section Liseola in this study and section Elegans in previous study, variation in the IGS appears to offer considerable potential to resolve intraspecific relationship as well as interspecies or intersection.

Molecular cloning and foreign gene expression of restriction endonuclease fragments of the Hc nuclear polyhedrosis virus DNA (Hc nuclear polyhedrosis virus DNA 제한효소절편의 molecular cloning 과 외래 유전자 발현)

  • Lee, Keun-Kwang
    • Journal of fish pathology
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    • v.8 no.1
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    • pp.31-36
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    • 1995
  • Hc nuclear polyhedrosis virus DNA genome was digested with EcoRI endonuclease, these partial fragments were recombined into the pUC8 plasmid vector and transformed in E. coli JM 83 cell. The genome DNA has 24 EcoRI fragments and 12 fragments of them were subcloned. The four recombinants were named as eNP-O, eNP-Q, eNP-R and eNP-S. The expression of foregin gene of the recombinants was investigated by analysing protein patterns on the SDS-PAGE. The eNP-O, eNP-Q and eNP-R were expressed a different weight of protein as comparision with potypeptide bands of E. coli JM 83 host cell.

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Morphology and Biochemcial Characteristics of a Nuclear Polyhedrosis Virus Isolated from the Oriental Tobacco Budworm, Helicoverpa assulta (Guenee) (담배나방 핵다각체병바이러스의 형태 및 생화학적 특성)

  • 진병래;박현우;우수동;김우진;김우진;박범석;강석권
    • Korean journal of applied entomology
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    • v.34 no.3
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    • pp.218-223
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    • 1995
  • A nuclear polyhedrosis virus isolated from the oriental tobacco budworm larvae, Helicoverpa assulta (Guenee) was characterized by electron microscopy, SDS-PAGE, restriction endonuclease analysis and cross infectivity. The shape of a polyhedron was $1.0\mu\textrm{m}$ in average with icosahdral outline, and the virus particle was $65nm\times300nm$ in average with rod-shape. The nuclear polyhedrosis virus was contained a single nucleocapsid within a viral envelope embedded in a polyhedron. The polyhedral protein was composed of a single polypeptide with a M.W. of 31 Kd. The genome size of the virus by restriction endonuclease analysis was about 120 Kb. Among several nuclear polyhedrosis viruses, the nuclear polyhedrosis virus from Helicoverpa assulta (HaNPV) and Autographa california nuclear polyhedrosis virus (AcNPV) were infected the oriental tobacco budworm larvae.

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16S rDNA-PCR and RFLP Analysis for rapid identification of Spoilage Bacteria from low Salt Cucumber Brine (저염 발효오이로부터 16S rDNA-PCR과 RFLP분석을 통한 부패균의 신속한 확인)

  • 김재호;장혜영
    • KSBB Journal
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    • v.19 no.1
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    • pp.72-77
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    • 2004
  • The aim of this study was to isolate and identify the spoilage bacteria in the low salt cucumber brine. The PCR amplicons comprising a portion of the 16S rRNA gene of the isolated colonies were directly sequenced and the untrimmed whole sequencing results of the unknown strains were aligned with the type strains using BLAST of NCBI. Then Sequence Aligner and Sequence Match of RDP confirmed the outcome. The identified isolates were eight species and belong to three genuses: Clostridium, Lactobacillus, and Bacillus. The RFLP pattern of the 16S rRNA gene of isolates verified the identified species. From now on the complex spoiling process of law salt fermented cucumber could be analyzed using the isolated species individually or with certain combinations.

Profile Analysis of Bacteria in Human Hands Using the Terminal Restriction Fragment Length Polymorphism (T-RFLP) Analysis (제한효소 절편길이 다형성(T-RFLP) 분석기법을 이용한 손에 서식하는 세균의 군집조성 분석)

  • Park, Jisun;Kim, Seung Bum
    • Journal of Science Criminal Investigation
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    • v.11 no.4
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    • pp.276-282
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    • 2017
  • As evidence that can be obtained at the crime scene, the importance of micro-evidences has been recognized in recent years with the development modern molecular-level analytical techniques. These micro-evidences include substances useful for personal identification such as DNA, but it is difficult to collect only the evidences showing individual characteristics every time at the crime scene. Therefore, development of new research approaches for the discovery and application of micro-evidence candidates is in increasing demand. For this purpose, skin microbial communities of bacteria inhabiting the palms of 16 people were collected and terminal-restriction enzyme fragment length polymorphism (T-RFLP) analysis was carried out to examine the potential for the application in personal identification. As a result, 16 different electropherograms were obtained, and various taxa including Staphylococcus and Bacillus were shown to produce different T-RF profiles among individuals. These results were analyzed with the factors affecting the microbiota such as sex and working environment of individuals.

Codon 311 Polymorphism of Paraoxonase-2 Gene and Hypertension in Korean (한국인에서 Paraoxonase-2 유전자의 Codon 311 다형성에 관한 연구)

  • Jang Dai Ho;Kang Byung Yong;Lee Kang Oh
    • Environmental Analysis Health and Toxicology
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    • v.19 no.2
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    • pp.135-140
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    • 2004
  • 고혈압에서 지질대사 이상은 빈번히 나타나는 현상으로, 지질대사 이상에 관여하는 유전자들은 고혈압의 발병원인을 규명하기 위한 후보 유전자로 인식되어 왔다. 이에 본 연구에서는 paraoxonase 2(PON2) 유전자에 존재하는 Cys311Ser다형성을 유전자 표지로 이용하여 한국인 집단에서 이 유전자 표지가 고혈압과 관련성이 있는 지를 조사하고자 하였다. 연구 대상은 총 195명으로, 이들 중에서 82명은 고혈압 환자 군이었고, 나머지 113명은 정상 혈압 군이었다. PON2 유전자의 Cys311Ser 다형성을 분석하기 위해서 중합효소 연쇄반응과 제한 효소인 Dde Ⅰ처리를 수행하여 유전자형을 결정하였다. 연구 결과, Cys/Ser이 형접합체를 갖는 사람들이 고혈압군에서 유의하게 높은 빈도로 나타났으며(P<0.05),다른 신체 계측치 및 혈청내 지질 농도와는 유의한 관련성을 나타내지 않았다. 본 연구에서 관찰된 이러한 관련성이 기능적인 연관인지 혹은 연관불평형에 의한 결과인지에 대해서는 보다 더 많은 연구 대상을 이용한 추시를 통해 밝혀질 수 있을 것으로 생각된다.

Isolation of Myxobacteria from Soil and RFLP Analysis of 16S rDNA Fragments. (토양으로부터 Myxobacteria의 분리 및 165 rDNA RFLP분석)

  • 김수광;최병현;김종균;이병규;강희일
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.187-191
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    • 2003
  • In an attempt to isolate myxobacteria from soil samples, we isolated swarm and fruiting body forming bacteria that have bacteriolytic activity on Coli-spot agar plate. For the classification of myxobacteria, 16S rDNA RFLP patterns were analyzed. Amplified 16S rDNAs of myxobacteria type strains (Family I, II, III and IV), negative control strains and soil-isolates were restricted with HaeIII, EcoRI and EcoRV, respectively. We found that the soil-isolates belongs to myxobacteria Family I, II, III.

Interstrain polymorphisms of isoenzyme profiles and mitochondrial DNA fingerprints among seven strains assigned to Acanthamoeba polyphaga (대식가시아메바(Acmthamoebapokphaga) 일곱 분리주간의 동위효소 profile과 Mitochondria DNA fingerprint의 다양성)

  • Gong, Hyeon-Hui;Park, Jun-Hyeong;Jeong, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.33 no.4
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    • pp.331-340
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    • 1995
  • Interstrain polymorphisms of isoenzyme profiles and mitochondrial (Mt) DNA fingerprints were observed among seven strains of Acnnthnmoeba isolated from different sources and morphologically assigned to A. polvphngn. Mt DNA ringerprints by eight restriction endonucleases (Bgl II, Sca I, Cla I, EcoR I, Xbo I, Kpn I, Sal I, and Sst I) revealed considerable interstrain polymorphisms . Isoenzyme profiles revealed considerable interstrain polymorphisms for acid phosphatase, lactate dehydrogenase, and glucose-6- phosphate dehydrogenase while those for glucose phosphate isomerase , leucine aminopeptidase , and malate dehydrogenase showed similarity Despite of the interstrain polymorphisms, the isoengyme profiles and Mt DNA fingerprints of the strain Ap were found to be identical with those of the strain .tones . Mt DNA fingerprinting was found to be highly applicable for the strain identification, characterization, and differentiation. Key words: Acanthnmoebn polyphcga, interstrain polymorphism, isoenzyme profiles , Mt DNA fingerprints, strain differentiation, strain identification.

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Identification of Meat Species Using Species-Specific PCR-RFLP Fingerprint of Mitochondrial 12S rRNA Gene (미토콘드리아 12S rRNA 유전자의 종 특이적 PCR-RFLP Fingerprint를 이용한 식육 원료의 판별)

  • Park, Jong-Keun;Shin, Ki-Hyun;Shin, Sung-Chul;Chung, Ku-Young;Chung, Eui-Ryong
    • Food Science of Animal Resources
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    • v.27 no.2
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    • pp.209-215
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    • 2007
  • In order to develop a sensitive and reliable method for the species-specific molecular markers, PCR-RFLP assay of the mitochondrial DNA(mt DNA) 12S rRNA gene was exploited for the identification of the origin of animal meat species including cattle, pig, sheep, goat, horse, deer, chicken, duck and turkey. A specific primer pairs were designed, based on the nucleotide sequences of mt 12S rRNA gene, for the amplification of the highly conserved region in the gene of the animal species using PCR-RFLP technique. mt DNA was isolated from meat samples followed by DNA amplification using PCR with the specific primers. PCR amplification produced an approximately 455 bp fragment in each of these animal meats. To distinguish pleat species, the PCR amplicons were digested with restriction endonucleases Tsp5091 and MboI, respectively, which generates distinct RFLP profiles. The DNA profiles digested with Tsp5091 allowed the clear discrimination in the mammalian meat species and the DNA profiles digested with MboI in poultry meat species. Therefore, the PCR-RFLP profiles of mt 12S rRNA gene could be very useful to identify the origin of the raw materials in the raw meats as well as the processed meat products.