• Title/Summary/Keyword: PAT

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Transformation and Expression of the PAT Gene in Arabidopsis Tryptophan Mutants

  • Lim, Seon-Hee;Kim, Young-Soon;Cheong, Hyeon-Sook
    • Journal of Plant Biology
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    • v.39 no.4
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    • pp.243-247
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    • 1996
  • Phosphoribosylanthranilate transferase (PAT) catalyzes the second step of the tryptophan biosynthetic pathway and is encoded by a single-copy gene that complements all the visible phenotypes of the tryptophan mutant (trp1-100) of Arabidopsis. The trp1-100 is blue fluorescent under UV light becuase it accumulates anthranilate. To obtain a plant with reduced PAT activity, PAT1 genes with several internal deletions in different promoter regions (pHS 101, pHS102, pHS104, pHS105, and pHS107) were induced into trp1-100 via Agrobacterium. Then, homozygous T3 plants were isolated and examined for blue fluorescence. Introduction of the PAT1 gene fusants results in the reversion of fluorescence phenotype except in the case of pHS105. These results prompted us to perform a parallel analysis of anthranilate synthase and PAT interms of the genetic complementation. A plant line carrying pHS105 gene fusant does not completely complement the blue fluorescence but it accumulates less anthranilate than trp1-100. The activity of PAT was reduced in the transgenic mutant as well. The plant carrying these constructs will add to the growing collection of molecular tools for the study of the indolic secondary metabolism.

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Molecular Approaches for Cloning of Important Higher Plant Genes (고등식물의 유용 유전자 크로닝을 위한 분자적 접근)

  • ;Ala
    • KSBB Journal
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    • v.10 no.1
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    • pp.89-96
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    • 1995
  • An Avabidofsis thaliana gene encoding phosphoribosyl anthranilate transferase is shown to be the gene that is defective in blue fluorescent trp 1 mutant plants. This gene, named PAT1, coding region is homologous to those for the phosphoribosyl anthranilate transferase from many microorganisms. This is due to a defect in tryptophan biosynthesis that leads to an accumulation of anthranilate, a fluorescent intermediate in the tryptophan pathway. PAT1 is a single-copy gene that complements all of the visible phenotypes of the different trp1 mutants. Experiments to determine the regulation of the PAT1 gene are in progress. The wild-type PAT1 promoter and several promoter deletions of PAT1 gene have been transformed into Arabidopsis tryptophan mutants. These constructs might identify promoter elements that control this patterns. We have isolated the homozygous lines in T3 seeds and analysed the protein levels using PAT antibody and PAT protein level increased two fold in pHSl07. Finally, the potential of using PAT1 as a selectable marker or visible reporter of gene expression is being explored.

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XPC-PAT Polymorphism in Korean Thyroid Papillary Carcinoma (한국인 갑상선 유두상암종 환자에서 XPC-PAT 유전자 다형)

  • Tae, Kyung;Lee, Keun-Young;Kim, Hee-Ok;Lee, Yong-Seop;Lee, Hyung-Seok;Ahn, You-Hern
    • Korean Journal of Head & Neck Oncology
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    • v.22 no.2
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    • pp.117-122
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    • 2006
  • Background and Objectives : Thyroid carcinoma is the sixth commonest cancer in Korea and the papillary carcinoma is the most common type(88%) of the malignant thyroid tumors. Bulky DNA adducts formed by the carcinogens are repaired by DNA repair process, but failure to repair this DNA damage can cause mutations in oncogenes and tumor suppressor genes resulting in tumor formation. The xeroderma pigmentosum group C(XPC) gene is essential for this repair procedure and the XPC-PolyAT(PAT) polymorphisms may alter DNA repair capacity(DRC) and genetic susceptibility to cancer. Subjects and Methods : In a case-control study of 113 Korean patients with pathologically diagnosed thyroid papillary carcinoma and 65 control subjects, we investigated the association between the three XPC-PAT gene polymorphisms and thyroid papillary cancer susceptibility. Results : The frequency of the variant XPC-PAT allele was lower in the cases(0.349) than in the controls (0.423), but the difference was not significant(p=0.140). Using logistic regression adjusting for age and sex, risk for thyroid papillary cancer was not increased in the XPC-PAT-/+ and XPC-PAT+/+ compared to XPCPAT-/-(adjusted overall odds ratio[95% confidence intervals;95%CI]=0.52[0.26-1.03] and 0.62 [0.22-1.75], respectively; trend test, p=0.167). Conclusion : There are no relationship between the XPC-PAT polymorphism and the risk of thyroid papillary carcinoma in Korean population. Based on our results, XPC-PAT polymorphism do not modulate genetic susceptibility to thyroid papillary cancer.

Genetic Transformation of Panax ginseng with Herbicide Resistant Gene (제초제 저항성 유전자에 의한 인삼의 형질전환)

  • 양계진
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.6
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    • pp.353-357
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    • 2001
  • Transformation of ginseng plants was achieved by biolistic system with cotyledon explants and callus using phosphinothricin acetyl-transferase (PAT) gene resisting to a herbicide of Bialaphos. The binary vector for transformation was constructed with disarmed Ti-plasmid and with double 355 promoter. The introduced NPT II and PAT genes of the transgenic ginseng plants were successfully identified by the PCR, and the survival test on the medium with basta. The transgenic ginseng plants were propagated using repetitive secondary embryogenesis. The transgenic ginseng plantlets had normal structures of roots and shoots, and dormant buds for new year sprouting. We transferred the transgenic plants to greenhouse and observed the continuing growth until a new year.

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Mutagenicity Studies of the Herbicide-resistance Phosphinotricin Acetyltransferase (PAT) (제초제저항성단백질 Phosphinotricin Acetyltransferase (PAT)의 유전독성시험)

  • Jeong, Mi-Hye;You, Are-Sun;Lee, Je-Bong;Shin, Jin-Sup;Kim, Jin-Hwa;Han, Jeung-Sul
    • The Korean Journal of Pesticide Science
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    • v.8 no.1
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    • pp.22-29
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    • 2004
  • To evaluate mutagenicity of Phosphinotricin Acetyltransferase(PAT) which is expressed by the glufosinate-resistance gene pat, in vitro reverse mutation test using Salmonella typhimurium, chromosome aberration test using chinese hamster lung(CHL) cells and in vivo micronucleus test of mice were performed. In the reverse mutation, the PAT did not induce mutagenicity in Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 with and without metabolic activation at $5000{\mu}g/plate$. In the chromosome aberration test, the results showed no incidence of increased structural and numerical chromosome aberrations at any doses tested(100, 10, $1{\mu}g/mL$). In micronucleus test, the ratio of micronuclei was measured in polychromatic erythrocytes of bone marrow of male ICR mice intraperitoneally administrated with PAT(1250, 625, and 313 mg/kg), the results showed no incidence of increased micronucleated polychromatic erythrocytes (MNPCE). These results indicate that PAT might not have mutagenic potential in vitro and vivo systems.

Quantitative Analysis of Phosphinothricin-N-acetyltransferase in Genetically Modified Herbicide Tolerant Pepper by an Enzyme-Linked Immunosorbent Assay

  • Shim, Youn-Young;Shin, Weon-Sun;Moon, Gi-Seong;Kim, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.681-684
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    • 2007
  • An immunoassay method was developed to quantitatively detect phosphinothricin-N-acetyltransferase (PAT) encoded by the Bialaphos resistance (bar) gene in genetically modified (GM) pepper. The histidine-tagged PAT was overexpressed in Escherichia coli M15 (pQE3l-bar) and efficiently purified by $Ni^{2+}$ affinity chromatography. A developed sandwich enzyme-linked immunosorbent assay (S-ELISA) method (detection limit: $0.01{\mu}g/ml$) was 100-fold more sensitive than a competitive indirect ELISA (CI-ELISA) method or Western blot analysis in detecting the recombinant PAT. In real sample tests, PAT in genetically modified herbicide-tolerant (GMHT) peppers was successfully quantified [$4.9{\pm}0.4{\mu}g/g$ of sample (n=6)] by the S-ELISA method. The S-ELISA method developed here could be applied to other GMHT crops and vegetables producing PAT.

The Biodegradability and Toxicity of Polyoxyethylene-Alkyl Citric Diester-Triethanolamine Detergent (Polyoxyethylene-Alkyl Citric Diester-Triethanolamine의 생분해 및 독성에 관한 조사 연구)

  • Kwon, Sook-Pyo;Chung, Yong
    • YAKHAK HOEJI
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    • v.23 no.1
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    • pp.1-6
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    • 1979
  • The authors investigated the biodegradability and toxicity of polyoxyethylene-alkyl citric diester-triethanolamine(PAT), which is one of new non-ionic detergents. 1) The PAT is very biodegradable, which 50% is biodegraded for one day and 100% for eight days by the activated sludge treatment and 40% for one day and 93% for eight days by the aerobic domestic sewage treatment. 2) $LD_{50}$ of PAT for chicken, mouse, and rat by oral adminstration are 7.1g/kg, 8.4g/kg and 14.0g/kg reactively. 3) $TL_{m}$ of PAT for goldfish is 64.0mg/l in 24bours. 4) No reaction for humun skin is determined by PAT.

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Developing a Performance Appraisal Tool for Neonatal Intensive Care Unit Registered Nurses (신생아중환자실 간호사 업무수행 평가도구 개발)

  • Park, Kwang-Ok;Lee, Yoon-Young
    • The Journal of Korean Academic Society of Nursing Education
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    • v.17 no.2
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    • pp.267-276
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    • 2011
  • Purpose: This study was done to develop a performance appraisal tool (PAT) for neonatal intensive care unit (NICU) registered nurses (RNs). Methods: The PAT was developed in three steps: 1) a standard of NICU nursing was established; 2) a draft was made; and 3) the PAT was ratified and its reliability and validity were tested. Results: The standard of practice of NICU nursing was predominantly based on role description and responsibilities for nurses. We identified 4 domains of nursing: professional practice, education, research, and leadership. The validity score for each item ranged from 3.93 to 3.27. The PAT consisted of 76 indicators, 49 reflecting components of professional practice, 5 related to responsibility for education, 3 representing research, and 19 relating to leadership. Cronbach's ${\alpha}$ averaged 0.99 for the 76 items. Conclusions: The PAT for NICU RNs that we created was found to be reliable and valid. This PAT would be very useful in evaluating nursing performance and facilitate the professional growth of nurses.

A Study on Antigencity (Immunotoxicity Study) to the Expressed Proteins of ${\beta}$-Carotene Biofortified Rice (베타카로틴강화미 발현단백질에 대한 항원성연구)

  • Park, Soo-Jin;Jeong, Mi-Hye;Chang, Hee-Seop;O, Jin-Cheol;Park, Kyung-Hun;Park, Jae-Yup
    • The Korean Journal of Pesticide Science
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    • v.15 no.3
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    • pp.289-297
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    • 2011
  • As part of safety evaluation of 2A (amono acid), PAT (phosphinotricin Acetyl-transferase), CtrI (Carotene desaturase) and PSY (phytoene synthase), the expressed proteins inserted to ${\beta}$-carotene Biofortified rice were tested for antigencity test. As a result, the group of administering high-concentration PAT, the expressed protein, showed a great content of total WBC; however, other expressed proteins did not show much difference. Against ASA (Active Systemic Anaphylaxis) test, the group of administering high-concentration PAT, the expressed protein, showed mild or medium degree of symptoms, but there was no dead entity. According to the result of the PCA (Passive Cutaneous Anaphylaxis test), the group of administering high-concentration PAT, 2A, PSY, and mixture of expressed proteins indicated positive response in low anti-serum concentration, and the group of administering the clinical concentration of mixture indicated mild positive response. However, because the group of administering the clinical concentration of expressed proteins, PAT, 2A, PSY, and CtrI, did not show positive response, it is thought that IgE is not generated. Further studies are needed to verify the safety of ${\beta}$-carotene Biofortified rice.

A Study on the Dielectric and the Piezoelectric Properties of xPb$(Al_{1/2}Ta_{1/2})O_3$-(1-x)Pb$(Zro_{0.52}Ti_{0.48})O_3$ Ceramics (xPb$(Al_{1/2}Ta_{1/2})O_3$-(1-x)Pb$(Zro_{0.52}Ti_{0.48})O_3$ 세라믹스의 유전 및 압전특성에 관한 연구)

  • Kang, Do-Won;Kim, Tae-Yoal;Oh, Jae-You;Oh, Eui-Kyun;Kim, Seok-Su;Kim, Beom-Jin;Kim, Myong-Ho;Park, Tae-Gone
    • Proceedings of the KIEE Conference
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    • 1999.07d
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    • pp.1726-1728
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    • 1999
  • Dielectric and piezoelectric properties of xPb$(Al_{1/2}Ta_{1/2})O_3$-(1-x)Pb$(Zro_{0.52}Ti_{0.48})O_3$ system were investigated. The highest density of $7.80g/cm^3$ for PAT-PZT ceramics of 5mol% PAT was obtained. The relative permittivity of PAT-PZT ceramics of 5mol% PAT was 1.642 at room temperature. The maximum value of electromechanical coupling factor $k_p$ of 55% and $k_t$ of 33% were obtained at the composition of 5mol% PAT. The grain sizes were reduced by increasing the amount of PAT, however mechanical quality factor$(Q_m)$ had a minimum value of 44 at the composition of 5mol% PAT.

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