• Title/Summary/Keyword: DNA polymerase I

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Expression of Cytochrome $b_{5}$ Retropseudogenes in Hunam Blood

  • Hwang, Mi-Sun;Alan W.Steggles;Yoo, Min
    • Biomedical Science Letters
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    • v.9 no.3
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    • pp.167-170
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    • 2003
  • Cytochrome $b_{5}$($b_{5}$) can be found in a variety of tissues and plays a role in the electron transfer pathways. Several retropseudogenes (numbered as I, II, II, IV, V) have been identified and well investigated for their structures. However, retropseudogene I is not clear in terms of its location on the chromosome. In addition the structure and the exression of retropseudogene V have not been confirmed. To examine the structure of bs retropseudogenes V and to see whether it is expressed in human blood we applied recombinant DNA technologies including polymerase chain reaction (PCR) and DNA sequencing. Retropseudogene V turned out to contain open reading frame (ORF) within its structure, however, no evidence of its expression was detected. Retropseudogene I was also found on the chromosome V. This study should contribute to the understanding of the structure of bs gene family.

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Phylogenetic Analysis by RFLP and Sequencing of Mitochondrial DNA in a Korean Population

  • Lee, Jin-Young;Kim, Heui-Soo;Ha, Bae-Jin;Park, Yeong-Hong
    • Archives of Pharmacal Research
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    • v.29 no.1
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    • pp.88-95
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    • 2006
  • Analysis of molecular nature of mitochondrial DNA (mtDNA) could be powerful marker for anthropological studies of modern populations. While population genetic studies on mtDNA have been reported for several ethnic groups, no such study has been documented for the Korean population. We surveyed mtDNA polymorphisms in the HVS I of noncoding D-loop region and its upstream region from 430 unrelated healthy Korean population by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct sequencing analysis. PCR product with 2,790 bp spanning the specific mtDNA region (mt13715-16504) was subjected to RFLP analysis using 6 restriction enzyme (Hinf I, Hae III, Alu I, Dde I, Mbo I, Rsa I). On the PAUP analysis of PCR-RFLP results, 38 mtDNA haplotypes (Hap 1-38) were detected in the Korean populations, which were classified into 11 haplogroups (Grp 1-11) of related haplotypes encompassing all 38 haplotypes. In comparison of sequencing data with Anderson's reference sequence, the transition type was more prevalent than the transversion type. Insertions or deletions were not found. In addition, three of the polymorphic sites (A16240C, A16351G, G16384A) in HVS-I region are determined newly. The polymorphic sites were distributed randomly in the region, though the frequency at each site was variable. Thus, this research might be required for the genealogical study of Orientals.

Application of 16S rDNA PCR-RFLP Analysis for the Rapid Identification of Weissella Species (Weissella 속 유산균의 빠른 동정을 위한 16S rDNA PCR-RFLP 분석법의 적용)

  • Lee, Myeong-Jae;Cho, Kyeung-Hee;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.455-460
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    • 2010
  • A polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis was applied to detect and identify ten Weissella spp. frequently found in kimchi. The previously reported genus-specific primers designed from 16S rDNA sequences of Weissella spp. were adopted but PCR was performed at the increased annealing temperature by $4^{\circ}C$. The sizes of amplified PCR products and restricted fragments produced by AluI, MseI, and BceAI endonucleases were well correspond with the expected sizes. W. kandleri, W. koreensis, W. confusa, W. minor, W. viridescens, W. cibaria, and W. soli were distinguished by AluI and MseI and W. hellenica and W. paramesenteroides were identified by BceAI. W. thailandensis was distinguished when restriction pattern of other species was compared but identified by the single use of MspI.

Identification and Differentiation of Cucumber Mosaic Virus Isolated from Forsythia koreana (CMV-Fk) Using PCR Techniques (PCR기법을 이용한 오이 모자이크 바이러스 개나리 분리주(CMV-Fk)의 동정과 구분)

  • 이상용;박선정;최장경
    • Korean Journal Plant Pathology
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    • v.14 no.4
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    • pp.308-313
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    • 1998
  • Reverse transcription and polymerase chain reaction (RT-PCR) techiniques were used to identification and differentiation of cucumber mosaic virus isolated from Forsythia koreana (CMV-Fk). RT-PCT used by two set of 20-mer primers one was CMV-common primers and another was CMV subgroup I-specific primers designed in a conserved region of the 3' end of CMV RNA3, amplified about 490 bp and 200 bp DNA fragments from CMV-Fk, respectively. CMV could be detected by RT-PCR at a dilution as low as 10-4 in forsythia crude sap extracts. Restriction enzyme analysis of RT-PCR products using EcoRI and MspI showed that CMV-Fk belonged to CMV subgroup I. But, analysis of RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR) showed heterogeneity of RNA3 between CMV-Fk and CMV-Y as a member of subgroup I.

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Genetic comparison between Spirometra erinacei and S. mansonoides using PCR-RFLP analysis (만손열두조충과 북미열두조충의 중합효소연쇄반응-마디길이여러꼴 분석법을 이용한 유전 형질 비교)

  • LEE, Soo-Ung;HUH, Sun;PHARES, C. Kirk
    • Parasites, Hosts and Diseases
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    • v.35 no.4
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    • pp.277-282
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    • 1997
  • The only observed morphological difference between Spirometra erinqsei and S. mcnsonoides is the uterine shape of the mature proglottid. Two species of worms are thought to be evolutionarily closely related. Biomolecular colnparison of the ho worms by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was conducted to observe the genetic distance. The 285 rDNA, mitochondrial cytochrome c oxidase subunit I (mCOI), and ribosomal internal transcribed spacer 1 (ITSI) fragments were obtained from the worms by PCR. The PCR products were cleaved by 5 four-base pair restriction enzyme combinations (Msp I, Hae III, Alu I, Cfo I, Rsa I) , electrophoresed and analyzed with PAUP 3.1.1. The fragment Patterns or 285 rDNA and Lni demonstrated that two worms were in identical systematic tree with bootstrap number 94 and 100, respectively As for mCOI, bootstrap number was 74 in a different tree. Above results are indicative of recent common ancestry between S. etinocei and S. mansonoides.

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Investigation of functional roles of transcription termination factor-1 (TTF-I) in HIV-1 replication

  • Park, Seong-Hyun;Yu, Kyung-Lee;Jung, Yu-Mi;Lee, Seong-Deok;Kim, Min-Jeong;You, Ji-Chang
    • BMB Reports
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    • v.51 no.7
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    • pp.338-343
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    • 2018
  • Transcription termination factor-1 (TTF-I) is an RNA polymerase 1-mediated transcription terminator and consisting of a C-terminal DNA-binding domain, central domain, and N-terminal regulatory domain. This protein binds to a so-called 'Sal box' composed of an 11-base pair motif. The interaction of TTF-I with the 'Sal box' is important for many cellular events, including efficient termination of RNA polymerase-1 activity involved in pre-rRNA synthesis and formation of a chromatin loop. To further understand the role of TTF-I in human immunodeficiency virus (HIV)-I virus production, we generated various TTF-I mutant forms. Through a series of studies of the over-expression of TTF-I and its derivatives along with co-transfection with either proviral DNA or HIV-I long terminal repeat (LTR)-driven reporter vectors, we determined that wild-type TTF-I downregulates HIV-I LTR activity and virus production, while the TTF-I Myb-like domain alone upregulated virus production, suggesting that wild-type TTF-I inhibits virus production and trans-activation of the LTR sequence; the Myb-like domain of TTF-I increased virus production and trans-activated LTR activity.

Sequence comparisons of 28S ribosomal DNA and mitochondrial cytochrome c oxidase subunit I of Metagonimus yokogawai, M. takahashii and M. miyatai

  • Lee, Soo-Ung;Huh, Sun;Sohn, Woon-Mok;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.42 no.3
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    • pp.129-135
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    • 2004
  • We compared the DNA sequences of the genus Metagonimus: M. yokogawai, M. takahashii, and M. miyatai. We obtained 288 D1 ribosomal DNA (rDNA) and mitochondrial cytochrome c oxidase subunit I (mtCOI) fragments from the adult worms by PCR, that were cloned and sequenced. Phylogenetic relationships inferred from the nucleotide sequences of the 28S D1 rDNA and mtCOI gene. M. takahashii and M. yokogawai are placed in the same clade supported by DNA sequence and phylogenie tree analysis in 28S D1 rDNA and mtCOI gene region. The above findings tell us that M. takahashii is closer to M. yokogawai than to M. miyatai genetically. This phylogenetic data also support the nomination of M. miyatai as a separate species.

Analysis of Mitochondrial DNA in Patients with Essential Tremor (본태성 수전증 환자의 미토콘드리아 DNA 분석)

  • Lee, Uhn;Yoo, Young Mi;Yoo, Chan Jong
    • Journal of Korean Neurosurgical Society
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    • v.29 no.2
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    • pp.188-195
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    • 2000
  • Objective : Essential tremor(ET) is the most common movement disorder, however, there has been little agreement in the neurologic literature regarding diagnostic criteria for ET. Familial ET is an autosomal dominant disorder presenting as an isolated postural tremor. The main feature of ET is postural tremor of the arms with later involvement of the head, voice, or legs. In previous studies, it was reported that ET susceptibility was inherited in an autosomal dominant inheritance. As previous results, it would suggest that ET might be associated with defect of mitochondrial or nuclear DNA. Recent studies are focusing on molecular genetic detection of movement disorders, such as essential tremor and restless legs syndrome. Moreover, authors have analysed mitochondrial DNA(mtDNA) from the blood cell of positive control(PC) and ET patients via long and accurate polymerase chain reaction(LA PCR). Materials & Methods : Blood samples were collected from PC and 9 ET patients. Total DNA was extracted twice with phenol followed by chloroform : isoamylalcohol. For the analysis of mtDNA, LA PCR was performed by mitochondrial specific primers. Results : With this technique, deletions of large quantities were detected within several regions of mtDNA in ET patients except for D-loop and CO I regions. Conclusion : The authors believe that ET is a genentic disorder with deficiency of mitochondrial DNA multicomplexes and mitochondiral dysfunction could be one of major causative factors of ET. Mitochondrial dysfunction may play an important role in the pathogenesis and possibility of disease progression among familial group with ET patients.

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Restriction Fragment Length Ploymorphism of PCR Amplified Ribosomal DNA Among Korean Isolates of Phytophthora

  • Hong, Seung-Beom;Jee, Hyeong-Jin;Lee, Seung-Im;Go, Seung-Joo
    • The Plant Pathology Journal
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    • v.15 no.4
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    • pp.228-235
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    • 1999
  • Genetic diversity of ninety-five Korean isolates of Phytophthora was investigated on the basis of PCR-RFLP of ribosomal DNA. The isolates were previously identified as following fifteen species by mycological and cultural characteristics; P. boehmeriae, P. cactorum, P. cambivora, P. capsici, P. cinnamoni, P. citricola, P. citrophthora, P. cryptogea, P. drechsleri, P. erythroseptica, P. infestans, P. megasperma, P. nicotianae, P. palmivora and P. sojae. The regions of small subunit (SSU) and internal transcribed spacer (ITS) of rDNA were amplified with primer pair, NS1 and ITS4, by polymerase chain reaction (PCR) and digested with nine restriction enzymes. P. boehmeriae, P. cactorum, P. cambivora, P. capsici, P. cinnamomi, P. citricola, P. citrphthora, P. infestans, P. nicotianae and P. palmivora showed specific band patterns for each species. However, P. sojae and P. erythroseptica presented identical band patterns and P. cryptogea, P. drechsleri and P. megasperma were divided into six groups, which were not compatible with delineation of the species. A group originated from cucurbits showed distinct band patterns from other groups, but the other five groups were closely related within 96.0% similarity, forming one complex group. Consequently, Korean isolates of Phytophthora were divided into thirteen genetic groups and each group was readily differentiated by comparing digestion patterns of AvaII, HaeIII, MboI, HhaI and MspI. Therefore, PCR-RFLP of rDNA using the five enzymes can be used to differentiate or identify the Phytophthora species reported in Korea so far.

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