• 제목/요약/키워드: TaqI

검색결과 88건 처리시간 0.022초

Construction of a Fusion-Stoffel Fragment to Improve 3′-5′Exonuclease Activity

  • CHOI, HYEJA;YOUNGSOO KIM
    • Journal of Microbiology and Biotechnology
    • /
    • 제8권6호
    • /
    • pp.669-675
    • /
    • 1998
  • Taq DNA polymerase exhibits a sizable drawback compared to the other thermophilic DNA polymerases in that it demonstrates lower proof-reading activity due to the deficiency of 3'-5'exonuclease activity. A study was undertaken to improve the 3'-5' exonuclease activity in the PCR of Taq DNA polymerase. The three-dimensional structural alignment of the polymerase and 3'-5' exonuclease domains from the pol I family DNA polymerases explains why Taq DNA polymerase has just a background level of 3'-5'exonuclease activity. A comparison indicated that the two polymerase domains are very similar in primary and tertiary conformations, even though Taq DNA polymerase carries a much shorter 3'-5'exonuclease domain than that of E. coli DNA polymerase I. Those two polymerase domains were interchanged between Taq DNA polymerase and E. coli DNA polymerase I. The 3'-5' exonuclease domain from E. coli DNA polymerase I was separated and pasted into the polymerase domain of Taq DNA polymerase I, which resulted in a functional fusion-Stoffel fragment. The 3'-5'exonuclease activity of the fusion-Stoffel fragment increased up to 48% of the value of the Klenow fragment, while that of Taq DNA polymerase remained at 6.0% of the Klenow fragment.

  • PDF

Construction of Two Metal-ion Binding Sites to Improve the 3′-5′Exonuclease Activity of Taq DNA Polymerase

  • Park, Yong-Hyun;Kim, Jong-Moon;Choi, Hye-Ja;Kim, Seog-K.;Kim, Young-Soo
    • Journal of Microbiology and Biotechnology
    • /
    • 제8권5호
    • /
    • pp.471-477
    • /
    • 1998
  • Taq DNA polymerase from Thermus aquaticus is very useful in the polymerase chain reaction. Taq DNA polymerase is classified in the pol I family, represented by E. coli DNA polymerase I. The three-dimensional structural alignment of 3'-5'exonuclease domains from the pol I family DNA polymerases explains why Taq DNA polymerase does not carry out proofreading in polymerase chain reactions. Three sequence motifs, Exo I, II, and III, must exist to carry out 3'-5'exonuclease activity for proof- reading by a 3'-5'exonuclease reaction, but these are abolished in Taq DNA polymerase. The key catalytic module in 3'-5'exonuclease is two metal ions chelated by four active-site carboxylic amino acids. Taq DNA polymerase was mutagenized to construct the catalytic module in the active site. The circular dichroism technique supported the formation of the catalytic module, and the radioactive assay showed that the 3'-5'exonuclease activity doubled in the mutant Taq DNA polymerase.

  • PDF

Staphylococcus aureus에서 분리된 plasmid상의 항생물질 저항성 인자에 관한 연구 : Chloramphenicol 저항성 인자의 클로닝 (Studies on antibiotics resistance gene in Staphylococcus aureun Plasmid: Cloning of chloramphenicol resistance determinant)

  • 권동현;김영선;변우현
    • 미생물학회지
    • /
    • 제24권4호
    • /
    • pp.341-351
    • /
    • 1986
  • 항생제에 대하여 다중 저항성을 갖는 Shaphylococcus aµreus D-H-1으로부터 chloram phenicol(Cm) 저항성 유전자를 가진것으로 확인된 R-plasmid(pSBK203, 2.5MdaJ.)를 분리하여 Bacillus subtilis BD 170 내에셔 발현시켰으며 이 Cm 저항성 유전자를 cloning하기 위하여 pSBK203상의 제한효소 인식부위를 결정하였다. pSBK203 상의 TaqI 부분 절편 0.3 kb을 pBD9 CZaI 인식부위내에 삽입하여 얻은 재조합 plasmid(pTQ16)와 TaqI부 분절편과 O.lkb 엇갈럼이 있는 RsaI 단일절떤(1. 3 kb} pBR322의 Seal 인식부위에 삽입하여 얻은 재조합 p plasmid(pHW20)에서 Cm 저항성이 획득되었다. pBD9 및 pBR322 상에 삽입된 두 절펀내에 Hinf, Taq I 빛 BglII의 제한효소 인식부위가 존재하였으며 이들 중 행III 인식부위에 의하여 Cm 저항성이 불활성 되었다.

  • PDF

Association Study between Vitamin D Receptor Gene Polymorphism and Adult Periodontitis in Korea

  • Kang, Byung-Yong;Ha, Nam-Joo
    • Animal cells and systems
    • /
    • 제7권2호
    • /
    • pp.145-149
    • /
    • 2003
  • Adult periodontitis is a chronic inflammatory disease whose etiology is not well defined. Recent studies have shown that vitamin D receptor gene has been a candidate for the susceptibility of adult periodontitis. The purpose of this study is to investigate the frequency of Taq I restriction fragment length polymorphism (RFLP) in the vitamin D receptor gene in nan periodontically healthy controls and 28 adult periodontitis patients. Taq I RFLP in the vitamin D receptor gene was detected by PCR amplification, followed by restriction enzyme digestion and 2% agarose gel electrophoresis. There was no significant difference in the distribution of Taq I RFLP between healthy controls and adult periodontitis group (P > 0.05). Thus, Taq I RFLP in the vitamin D receptor gene may not confer the susceptibility to adult periodontitis in Korean population. However, t allele distributions of this RFLP showed various frequencies among ethnic groups studied. Further studies in other ethnic groups will be required.

Central Asian Textile Motifs in Late Sasanian Art: On the Origin of Some Decorative Elements at Taq-i Bustan

  • COMPARETI, Matteo
    • Acta Via Serica
    • /
    • 제6권2호
    • /
    • pp.21-36
    • /
    • 2021
  • This paper discusses textile motifs depicted in the hunting panels inside the late Sasanian larger grotto at Taq-i Bustan. Scholars of Iranian art have referred to these rock reliefs in order to trace the origin of Sasanian textile decorations and their exportation in the whole medieval Eurasian continent. This however does not seem to be the case. In fact, the textile motifs reproduced at Taq-i Bustan could be a good term of comparison only for late Sasanian textile production. Moreover, nothing like this has appeared in previous Sasanian rock reliefs. For this reason, the present paper argues that some of those motifs could actually be importations from Central Asia where reproductions of textiles embellished with those motifs were very popular. Islamic written sources on Taq-i Bustan rock reliefs could be very useful to support some ideas expressed in this paper.

정신분열병환자에서 Taq I A 도파민 $D_2$ 수용체 다형성과 항정신병약물의 치료반응과의 연관 (The Relationship between Taq I A Dopamine $D_2$ Receptor Polymorphism and Therapeutic Response to Antipsychotics in Schizophrenic Patients)

  • 강철중
    • 생물정신의학
    • /
    • 제12권2호
    • /
    • pp.107-113
    • /
    • 2005
  • Purpose:In an attempt to predict the interpersonal differences of therapeutic response to antipsychotic drugs on pharmaco-genetic bases, this study was designed to investigate the relationship between the therapeutic response to antipsychotic drugs and Taq I A dopamine $D_2$ receptor polymorphism in schizophrenic patients. Methods:The subjects were 158 patients diagnosed with schizophrenia(DSM-IV). The therapeutic response to antipsychotic drugs was evaluated using the Treatment Response Scale(TRS) retrospectively. Patients were divided into two groups, dopamine receptor antagonist responders, and serotonin-dopamine antagonist responders. The patients' Taq I A dopamine $D_2$ receptor polymorphism was determined by polymerase chain reaction(PCR) and restriction fragment length polymorphism(RFLP). Results:The dopamine receptor antagonist responders had the A1 allele in significantly higher incidences (${\chi}^2$(1)=4.875, p=0.027, two-tailed). No significant difference was found among the serotonin-dopamine antagonist responders between those with or without the A1 allele. Conclusions:The patients with the A1 allele responded better to dopamine receptor antagonists than those with no A1 allele. Based on these results, it is suggested that the pharmacological effect of dopamine receptor antagonists can be predicted depending on the presence of the A1 allele in schizophrenic patients.

  • PDF

East-West Exchange of Costume Culture: Focusing on the Analysis of Taq-i Bustan Reliefs of the Sassanian Dynasty of Persia

  • CHANG, Youngsoo
    • Acta Via Serica
    • /
    • 제6권2호
    • /
    • pp.1-20
    • /
    • 2021
  • The Taq-i Bustan reliefs are representative works from the Sassanian dynasty of the 4th to 7th centuries. This study analyzes the costumes depicted in the Taq-i Bustan reliefs to gain understanding of the phenomena of cultural exchange between the East and West by observing the foreign cultural elements appearing in the Sassanian costumes of that time. Literature study and artifacts analysis were conducted in parallel. External elements appearing in Taq-i Bustan's costume were Greek-Roman and Central Asian. The tunics and trousers of the gods and the trousers of kings (Ardashir II, Shapur II and Shapur III) were made of thin fabric and showed many wrinkles, a characteristic of Greek and Roman clothing. On the spandrel above the arch of the great grotto of Khusrau II are depicted the goddesses of Victory, in a Greco-Bactrian style. Among the costume elements of Taq-i Bustan, there were also Central Asian elements observed. One Central Asian costume element was the round clasp ornament for tying the trousers. The side slits and hem of the tunic were presented in the style of the Sogd clothing of Central Asia in the 6th and 7th centuries, while the pearl rounded pattern was activated in Sogd, Kucha and Kizyl in the 7th and 8th centuries. These reliefs are considered important evidence of eastern influences in Sassanian culture.

도파민 D2 수용체 다형성과 보상의존성 성격특성과의 관련성 (Association between D2 Dopamine Receptor Gene Polymorphisms and Reward Dependence Personality Traits)

  • 강이헌;이헌정;곽강호;김린;이민수;서광윤
    • 생물정신의학
    • /
    • 제11권1호
    • /
    • pp.26-32
    • /
    • 2004
  • Background:The dopaminergic genes have been implicated with some personality traits. Many recent studies indicated that there is a correlation between D2 dopamine receptor gene(DRD2) polymorphisms and the personality traits. The purpose of this study is to investigate a possible association between DRD2 gene (TaqI A, TaqI B) polymorphism and personality traits. Methods:The subjects were consisted of 173 blood-unrelated young female Koreans with a mean age(${\pm}SD$) of 13.88(${\pm}0.29$) years. These volunteers were recruited from one of the junior high schools in Seoul and were tested by the Korean version of the Temperament and Character Inventory(TCI). Genotyping of the DRD2 polymorphisms by PCR methods were carried out. Two DRD2 gene polymorphisms were classified and individually assessed as follows:TaqI A1+ vs A1-, TaqI B1+ vs B-. The associations between the TCI scores and TaqI A, TaqI B polymorphisms were assessed by Student's t-test. Results:In the 173 subjects, the allele frequencies of the DRD2 TaqI A1, TaqI B1 alleles ranged from 0.42 to 0.43, and these results are quite different from the ranges of 0.15-0.20 in the case of a Caucasian population. The genotype frequencies of DRD2(TaqI A1, TaqI B1) variants showed no significant deviation from the Hardy-Weinberg equilibrium. RD4(dependence vs. independence) of Cloninger's TCI, a sub-dimension of Reward Dependence, was significantly higher in the subjects having DRD2 less frequent alleles than those without these alleles. Conclusion:This study suggests that the female subjects carrying the less frequent DRD2 alleles exhibited higher reward-dependent personality trait compared to those without these alleles.

  • PDF

한국인 사회공포증 환자에서 도파민 D2 수용체 TaqI A 유전자 다형성 : 예비적 연구 (Dopamine D2 Receptor Gene TaqI A Polymorphism in Korean Social Phobia Patients : Preliminary Study)

  • 김범조;임세원;신동원;오강섭;이민수
    • 생물정신의학
    • /
    • 제14권2호
    • /
    • pp.106-114
    • /
    • 2007
  • Objective : It is suggested that disturbance of dopaminergic system might be related to the possible mechanism of social phobia. The aim of this study was to investigate the possible association of DRD2 TaqI polymorphism and social phobia. Method : Fifty-one patients with social phobia and 200 comparison subjects were tested for DRD2 TaqI A polymorphism. The severity of social phobic symptoms was measured by self-report version of the Liebowitz Social Anxiety Scale(LSAS-SR) and Hamilton anxiety scale(HAM-A). Results : There was no signigicant difference in the genotype, allele frequency, A1 carrier frequency, and heterozygote frequency DRD2 TaqI A polymorphism between the social phobia patients and the control groups. However, we found significant decrease in somatic anxiety of the HAM-A in the patients having A2A2 homozygotes(p=0.014). In addition, patients having A1A2 heterozygotes showed more anxiety in two subscales (p=0.042 in anxiety, p=0.019 in performance) of the LSAS-SR. Conclusion : These results suggest that DRD2 A2 homozygote might have a protective role against somatic anxiety, and molecular heterosis of DRD2 TaqI A polymorphism might be related with more severe anxiety in social phobia.

  • PDF

사과 바이러스 검정을 위한 SYBR Green I 및 TaqMan probe 기반의 real-time PCR 검사법 개발 (Development of Real-time Quantitative PCR Assay based on SYBR Green I and TaqMan Probe for Detection of Apple Viruses)

  • 허성;정용석
    • 한국작물학회지
    • /
    • 제65권4호
    • /
    • pp.496-507
    • /
    • 2020
  • 본 연구는 사과 바이러스 ASGV, ASPV 및 ApMV를 각각 정밀하게 진단하고자 SYBR Green I 및 TaqMan probe, 두 종류의 다른 chemical dye를 사용하여 quantitative real-time PCR 검정법을 개발하고자 하였다. 1. 사과 바이러스 ASGV, ASPV 및 ApMV의 국내분리주를 바탕으로 하여 cloning 및 in vitro transcription을 수행해 10배 희석단위 표준시료를 제작하였다. 각 바이러스에 대한 SYBR Green I용 프라이머와 TaqMan probe용 프라이머 및 프로브를 디자인하였다. 2. 상기 제작된 프라이머와 프로브를 이용해 표준시료를 대상으로 real-time PCR을 수행하여 각 바이러스의 증폭곡선과 검량선을 구할 수 있었다. Real-time PCR 결과, SYBR Green I기반의 검정법은 TaqMan probe기반의 검정법 못지 않은 결과를 보여주었으며, 적은 비용에 대량 검정이 요구되는 곳에 효과적으로 응용될 수 있을 것이다. 3. 현장평가를 본 실험에서 개발된 TaqMan probe기반의 real-time PCR검정법과 기존의 RT-PCR검정법과 비교분석하였다. 그 결과 real-time PCR 검정법은 singleplex 및 multiplex RT-PCR보다 더 민감하고 정확한 결과를 내어 RT-PCR로 검출할 수 없는 농도까지 검정할 수 있음을 입증하였다. 4. 본 실험에서 개발한 real-time PCR검정법은 검역현장과 같은 대량의 검사가 요구되는 곳에서는 SYBR Green I 기반의 검정법을, 바이러스 연구분야에서는 세밀한 정량이 가능한 TaqMan probe 방식의 검정법이 활용될 것으로 기대한다.