• Title, Summary, Keyword: Marker residue

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Harmonization of MRL Setting for Compounds Used Both as Pesticides and as Veterinary Drugs with Regulatory Aspects - Cypermethrin in Food of Animal Origin (농약 및 동물용의약품으로 사용되는 약제의 잔류허용기준 설정 개선 - 축산물 중 cypermethrin의 잔류 사례)

  • Kwon, Jin-Wook
    • Korean Journal of Environmental Agriculture
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    • v.30 no.1
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    • pp.89-97
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    • 2011
  • BACKGROUND: Cypermethrins, possess eight isomers, used both as pesticide and as veterinary drug, were set different MRLs for livestock by CCPR and CCRVDF of Codex Alimentarius. Korea Food Code designates MRLs for livestock only as pesticide. METHODS AND RESULTS: This study presented necessaries of harmonization of MRL setting for compounds used both as pesticides and as veterinary drugs with regulatory aspects, showing an example of cypermethrin residue in livestock. CONCLUSION(S): For harmonization, following factors must be considered and recommended; designation of marker residue; alpha-cypermethrin, zeta- cypermethrin, and cypermethrin, clarification of the definition of target tissues; meat, fat, muscle, by-product, eggs, milk, and etc., method of analysis; clarification of target analytes of isomers, quantitation and calculation method as a principle of residue analysis.

Molecular discrimination of Panax ginseng cultivar K-1 using pathogenesis-related protein 5 gene

  • Wang, Hongtao;Xu, Fengjiao;Wang, Xinqi;Kwon, Woo-Saeng;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.43 no.3
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    • pp.482-487
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    • 2019
  • Background: The mixed-cultivation of different Panax ginseng cultivars can cause adverse effects on stability of yield and quality. K-1 is a superior cultivar with good root shape and stronger disease resistance. DNA markers mined from functional genes are clearly desirable for K-1, as they may associate with major traits and can be used for marker-assisted selection to maintain the high quality of Korean ginseng. Methods: Five genes encoding pathogenesis-related (PR) proteins of P. ginseng were amplified and compared for polymorphism mining. Primary, secondary, and tertiary structures of PR5 protein were analyzed by ExPASy-ProtParam, PSSpred, and I-TASSER methods, respectively. A coding single nucleotide polymorphism (SNP)-based specific primer was designed for K-1 by introducing a destabilizing mismatch within the 3' end. Allele-specific polymerase chain reaction (PCR) and real-time allele-specific PCR assays were conducted for molecular discrimination of K-1 from other cultivars and landraces. Results: A coding SNP leading to the modification of amino acid residue from aspartic acid to asparagine was exploited in PR5 gene of K-1 cultivar. Bioinformatics analysis showed that the modification of amino acid residue changed the secondary and tertiary structures of the PR5 protein. Primer KSR was designed for specific discrimination of K-1 from other ginseng cultivars and landraces. The developed real-time allele-specific PCR assay enabled easier automation and accurate genotyping of K-1 from a large number of ginseng samples. Conclusion: The SNP marker and the developed real-time allele-specific PCR assay will be useful not only for marker-assisted selection of K-1 cultivar but also for quality control in breeding and seed programs of P. ginseng.

Fingerprint of Marker Substances in Gami-Honghwa-Tang(KH-19) by HPLC-DAD (High Performance Liquid Chromatography-Diode Array Detector(HPLC-DAD)에 의한 가미홍화탕 (KH-19)의 지문 분석)

  • Yu Young-Beob;Yoon Yoo-Sik;Cho Gi-Ho
    • The Journal of Korean Medicine
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    • v.25 no.3
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    • pp.45-54
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    • 2004
  • Objectives : This study was aimed to evaluate marker substances in Gami-Honghwa-Tang (KH-19) by high performance liquid chromatography-photodiode array detector (HPLC-DAD). Gami-Honghwa-Tang is composed of nine crude herbs, Rehmanniae Radix Preparata, Angelicae Gigantis Radix, Cnidii Rhizoma, Paeoniae Radix, Corni Fructus, Moutan Cortex Radicis, Lycii Fructus, Carthami Flos and Glycyrrhizae Radix. Methods : The separation was performed on an Aquasil C18 (4.6×250mm) column by gradient elution with 0.05% TFA in H2O - 0.05% TFA in acetonitrile (0 min 100:0, 20 min 90:10, 40 min 70:30, 60 min 50:50, 80 min 0:100, 90 min 100:0) as the mobile phase at a flow-rate of 1.0 ml/min with detection at 190-800nm. Also we examined the contents for bacteria, pesticide residue and harmful heavy metals. Results : HPLC-DAD was employed to determine the quantities and the qualities of several marker substances such as 5­hydroxymethyl-2-furaldehyde (5-HMF), paeonol, loganin, paeoniflorin, glycyrrhizin, and decursin in the KH-19. There were no bacterial contents, pesticide residues, or harmful heavy metals. Conclusions : We suggest these results could be a useful evidence for quality control of KH-19. This method permits fingerprints of selected individual marker substances from herbal prescriptions without derivatization, multiple purification steps, or lengthy separation times.

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Study for the Standardization and Comparison by Processed Morindae Radix (파극천(巴戟天) 포제방법(?製方法)에 따른 품질표준화 연구)

  • Lee, Hye-Won;Chun, Jin-Mi;Lee, A-Yeong;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.20 no.4
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    • pp.133-140
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    • 2005
  • Objectives : We have been used many herbal medicines after processing to improve the effect, decrease toxicity and side-effect, and change property. We have studied the physico-chemical change and HPLC pattern of Morindae Radix by means of processing method. Methods : This study was investigated the contents of loss on drying, residue on ignition, residue on acid insoluble ignition, 50% ethanol extract and HPLC pattern of Morindae Radix(Morinda officinalis How.) by processed and non-processed. We have conducted Morindae Radix and Damnacanthi Radix which is circulated in herbal medicine market by forgery. Processed Morindae Radix was prepared by heating of added to salt(SP), liquor(LP) and Glycyrrhizae Radix solution(GP) for 20-40 minutes. Results and Conclusions : From this analysis, we found that the content of 50% ethanol extract was increased by processing method. And we were detected distinguishable marker of processed and non-processed from Morindae Radix(Morinda officinalis How.) by HPLC pattern analysis.

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Isolation of the Phosphoribosyl Anthranilate Isomerase Gene (TRP1) from Starch-Utilizing Yeast Saccharomycopsis fibuligera

  • Park, Eun-Hee;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.8
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    • pp.1324-1327
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    • 2015
  • The nucleotide sequence of the TRP1 gene encoding phosphoribosyl anthranilate isomerase in yeast Saccharomycopsis fibuligera was determined by degenerate polymerase chain reaction and genome walking. Sequence analysis revealed the presence of an uninterrupted open-reading frame of 759 bp, including the stop codon, encoding a 252 amino acid residue. The deduced amino acid sequence of Trp1 in S. fibuligera was 43.5% homologous to that of Komagataella pastoris. The cloned TRP1 gene (SfTRP1) complemented the trp1 mutation in Saccharomyces cerevisiae, suggesting that it encodes a functional TRP1 in S. fibuligera. A new auxotrophic marker to engineer starch-degrading yeast S. fibuligera is now available. The GenBank Accession No. for SfTRP1 is KR078268.

Molecular Genetics and Diagnostic Approach of Mucolipidosis II/III

  • Sohn, Young Bae
    • Journal of mucopolysaccharidosis and rare diseases
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    • v.2 no.1
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    • pp.13-16
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    • 2016
  • Mucolipidosis (ML) II/III are autosomal recessive diseases caused by deficiency of post-translational modification of lysosomal enzymes. The mannose-6-phosphate (M6P) residue in lysosomal enzymes synthesized by N-acetylglucosamine 1-phosphotransferase (GlcNAc-phosphotransferase) serves as recognition marker for trafficking in lysosomes. GlcNAc-phosphotransferase is encoded by GNPTAB and GNPTG. Mutations in GNPTAB cause severe ML II alpha/beta and the attenuated ML III alpha/beta. Whereas mutations in GNPTG cause the ML III gamma, the attenuated type of ML III variant. For the diagnostic approaches, increased urinary oligosaccharides excretion could be a screening test in clinically suspicious patients. To confirm the diagnosis, instead of measuring the activity of GlcNAc phosphotransferase, measuring the enzymatic activities of different lysosomal hydrolases are useful for diagnosis. The activities of several lysosomal hydrolases are decreased in fibroblasts but increased in serum of the patients. In addition, the sequence analysis of causative gene is warranted. Therefore, the confirmatory diagnosis requires a combination of clinical evaluation, biochemical and molecular genetic testing. ML II/III show complex disease manifestations with lysosomal storage as the prime cellular defect that initiates consequential organic dysfunctions. As there are no specific therapy for ML to date, understanding the molecular pathogenesis can contribute to develop new therapeutic approaches ultimately.

Isolation and functional analysis of three microsomal delta-12 fatty acid desaturase genes from Camelina sativa (L.) cv. CAME (카멜리나 (Camelina sativa L. cv. CAME)로부터 3 microsomal delta-12 fatty acid desaturase 유전자들의 분리 및 기능 분석)

  • Kim, Hyojin;Go, Young Sam;Kim, Augustine Yonghwi;Lee, Sanghyeob;Kim, Kyung-Nam;Lee, Geung-Joo;Kim, Gi-Jun;Suh, Mi Chung
    • Journal of Plant Biotechnology
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    • v.41 no.3
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    • pp.146-158
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    • 2014
  • Camelina sativa that belongs to Brassicaceae family is an emerging oilseed crop. Camelina seeds contain approximately 40% storage oils per seed dry weight, which are useful for human and animal diets and industrial applications. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. In this study, three CsFAD2 genes (CsFAD2-1, CsFAD2-2 and CsFAD2-3.1) were isolated from developing seeds of Camelina sativa (L.) cv. CAME. The nucleotide and deduced amino acid sequences of three CsFAD2 genes were compared with those from dicotyledon and monocotyledon plants including Camelina cultivars Sunesone and SRS933. Three histidine motifs (HECGH, HRRHH, and HVAHH) required for FAD activity and a hydrophobic valine or isoleucine residue, which is a SNP (single nucleotide polymorphism) marker related with enzyme activity are well conserved in three CsFAD2s. The expressions of CsFAD2-1 and CsFAD2-3.1 were ubiquitously detected in various Camelina organs, whereas the CsFAD2-2 transcripts were predominantly detected in flowers and developing seeds. The contents of oleic acids decreased, whereas the amounts of linoleic acid increased in dry seeds of transgenic fad2-2 lines expressing each CsFAD2 gene compared with fad2-2 mutant, indicating that three CsFAD2 genes are functionally active. The isolated CsFAD2 genes might be applicable in metabolic engineering of storage oils with high oleic acids in oilseed crops.