• Title/Summary/Keyword: Taq I

Search Result 88, Processing Time 0.111 seconds

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

  • CHOI, HYEJA;YOUNGSOO KIM
    • Journal of Microbiology and Biotechnology
    • /
    • v.8 no.6
    • /
    • 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
    • /
    • v.8 no.5
    • /
    • 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

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

  • 권동현;김영선;변우현
    • Korean Journal of Microbiology
    • /
    • v.24 no.4
    • /
    • pp.341-351
    • /
    • 1986
  • R-plasmid(pSBK203, 2.5Mdal) conferring chloramphenicol resistance was isolated from mutiple antibiotic resistant Staphylococcus aureus D-H-1. Bacillus subtilis BD170 was transformed by this plasmid and restriction enzyme clevage sites of this plasmid were mapped for the cloning of chloramphenicol resistance gene. Taq I partial digested fragment of pSBK203(1.3kb) inserted into Cla I site of pBD9 appears to have both regulatory region for induction and structural gene for chloramphenicol resistance whereas Rsa I fragment (1.3kb, both ends are staggered away 0.1Kb from those of Taq I fragment) inserted into Sca I site of pBR322 showed constitutive expression in E. coli. Hinf I, Taq I, and Bgl II restriction enzyme recognition sites are found in both Rsa I fragment and Taq I fragment. Among these, Bgl II recognition site was associated with chloramphenicol resistance.

  • PDF

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

  • Kang, Byung-Yong;Ha, Nam-Joo
    • Animal cells and systems
    • /
    • v.7 no.2
    • /
    • 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
    • /
    • v.6 no.2
    • /
    • 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.

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

  • Kang, Cheol Joong
    • Korean Journal of Biological Psychiatry
    • /
    • v.12 no.2
    • /
    • 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
    • /
    • v.6 no.2
    • /
    • 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.

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

  • Kang, Rhee-Hun;Lee, Heon-Jeong;Kwak, Kang-Ho;Kim, Leen;Lee, Min-Soo;Suh, Kwang-Yoon
    • Korean Journal of Biological Psychiatry
    • /
    • v.11 no.1
    • /
    • 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

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

  • Kim, Bum-Jo;Lim, Se-Won;Shin, Dong-Won;Oh, Kang-Seob;Lee, Min-Soo
    • Korean Journal of Biological Psychiatry
    • /
    • v.14 no.2
    • /
    • 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

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

  • Heo, Seong;Chung, Yong Suk
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.65 no.4
    • /
    • pp.496-507
    • /
    • 2020
  • Virus infections of apples result in lowered commercial qualities such as low sugar content, weakened tree vigor, and malformed fruits. An effective way to control viruses is to produce virus-free plants based on the development of an accurate and sensitive diagnostic method. In this study, real-time PCR assays based on SYBR Green I and TaqMan probes were developed for detecting ASGV, ASPV, and ApMV viruses. These methods can detect and quantify 103 to 1011 RNA copies/μL of each virus separately. Compared with methods with two different dyes, the SYBR Green I-based method was efficient for virus detection as well as for assay using the TaqMan probe. Field tests demonstrated that real-time PCR methods developed in this study were applicable to high-throughput diagnoses for virus research and plant quarantine.