• Title/Summary/Keyword: partial identification

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Identification of mountain-cultivated ginseng and cultivated ginseng using UPLC/oa-TOF MSE with a multivariate statistical sample-profiling strategy

  • Xu, Xin-fang;Cheng, Xian-long;Lin, Qing-hua;Li, Sha-sha;Jia, Zhe;Han, Ting;Lin, Rui-chao;Wang, Dan;Wei, Feng;Li, Xiang-ri
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.344-350
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    • 2016
  • Background: Mountain-cultivated ginseng (MCG) and cultivated ginseng (CG) both belong to Panax ginseng and have similar ingredients. However, their pharmacological activities are different due to their significantly different growth environments. Methods: An ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS)-based approach was developed to distinguish MCG and CG. Multivariate statistical methods, such as principal component analysis and supervised orthogonal partial-least-squares discrimination analysis were used to select the influential components. Results: Under optimized UPLC-QTOF-MS/MS conditions, 40 ginsenosides in both MCG and CG were unambiguously identified and tentatively assigned. The results showed that the characteristic components of CG and MCG included ginsenoside Ra3/isomer, gypenoside XVII, quinquenoside R1, ginsenoside Ra7, notoginsenoside Fe, ginsenoside Ra2, ginsenoside Rs6/Rs7, malonyl ginsenoside Rc, malonyl ginsenoside Rb1, malonyl ginsenoside Rb2, palmitoleic acid, and ethyl linoleate. The malony ginsenosides are abundant in CG, but higher levels of the minor ginsenosides were detected in MCG. Conclusion: This is the first time that the differences between CG and MCG have been observed systematically at the chemical level. Our results suggested that using the identified characteristic components as chemical markers to identify different ginseng products is effective and viable.

Isolation and Identification of Fungi from a Meju Contaminated with Aflatoxins

  • Jung, Yu Jung;Chung, Soo Hyun;Lee, Hyo Ku;Chun, Hyang Sook;Hong, Seung Beom
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1740-1748
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    • 2012
  • A home-made meju sample contaminated naturally with aflatoxins was used for isolation of fungal strains. Overall, 230 fungal isolates were obtained on dichloran rosebengal chloramphenicol (DRBC) and dichloran 18% glycerol (DG18) agar plates. Morphological characteristics and molecular analysis of a partial ${\beta}$-tubulin gene and the internal transcribed spacer (ITS) of rDNA were used for the identification of the isolates. The fungal isolates were divided into 7 genera: Aspergillus, Eurotium, Penicillium, Eupenicillium, Mucor, Lichtheimia, and Curvularia. Three strains from 56 isolates of the A. oryzae/flavus group were found to be aflatoxigenic A. flavus, by the presence of the aflatoxin biosynthesis genes and confirmatory aflatoxin production by high-performance liquid chromatography (HPLC). The predominant isolate from DRBC plates was A. oryzae (42 strains, 36.2%), whereas that from DG18 was A. candidus (61 strains, 53.5%). Out of the 230 isolates, the most common species was A. candidus (34.3%) followed by A. oryzae (22.2%), Mucor circinelloides (13.0%), P. polonicum (10.0%), A. tubingensis (4.8%), and L. ramosa (3.5%). A. flavus and E. chevalieri presented occurrence levels of 2.2%, respectively. The remaining isolates of A. unguis, P. oxalicum, Eupenicillium cinnamopurpureum, A. acidus, E. rubrum, P. chrysogenum, M. racemosus, and C. inaequalis had lower occurrence levels of < 2.0%.

Application of Molecular Methods for the Identification of Acetic Acid Bacteria Isolated from Blueberries and Citrus Fruits

  • Gerard, Liliana Mabel;Davies, Cristina Veronica;Solda, Carina Alejandra;Corrado, Maria Belen;Fernandez, Maria Veronica
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.193-204
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    • 2020
  • Sixteen acetic acid bacteria (AAB) were isolated from blueberries and citric fruits of the Salto Grande region (Concordia, Entre Rios, Argentina) using enrichment techniques and plate isolation. Enrichment broths containing ethanol and acetic acid enabled maximum AAB recovery, since these components promote their growth. Biochemical tests allowed classification of the bacteria at genus level. PCR-RFLP of the 16S rRNA and PCR-RFLP of the 16S-23S rRNA intergenic spacer allowed further classification at the species level; this required treatment of the amplified products of 16S and 16S-23S ITS ribosomal genes with the following restriction enzymes: AluI, RsaI, HaeIII, MspI, TaqI, CfoI, and Tru9I. C7, C8, A80, A160, and A180 isolates were identified as Gluconobacter frateurii; C1, C2, C3, C4, C5, C6, A70, and A210 isolates as Acetobacter pasteurianus; A50 and A140 isolates as Acetobacter tropicalis; and C9 isolate as Acetobacter syzygii. The bacteria identified by 16S rRNA PCR-RFLP were validated by 16S-23S PCR-RFLP; however, the C1 isolate showed different restriction patterns during identification and validation. Partial sequencing of the 16S gene resolved the discrepancy.

Isolation and identification of mammalian orthoreovirus type 3 from a Korean roe deer (Capreolus pygargus)

  • Yang, Dong-Kun;An, Sungjun;Park, Yeseul;Yoo, Jae Young;Park, Yu-Ri;Park, Jungwon;Kim, Jong-Taek;Ahn, Sangjin;Hyun, Bang-Hun
    • Korean Journal of Veterinary Research
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    • v.61 no.2
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    • pp.13.1-13.8
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    • 2021
  • Mammalian reovirus (MRV) causes respiratory and intestinal disease in mammals. Although MRV isolates have been reported to circulate in several animals, there are no reports on Korean MRV isolates from wildlife. We investigated the biological and molecular characteristics of Korean MRV isolates based on the nucleotide sequence of the segment 1 gene. In total, 144 swabs from wild animals were prepared for virus isolation. Based on virus isolation with specific cytopathic effects, indirect fluorescence assays, electron microscopy, and reverse transcription-polymerase chain reaction, only one isolate was confirmed to be MRV from a Korean roe deer (Capreolus pygargus). The isolate exhibited a hemagglutination activity level of 16 units with pig erythrocytes and had a maximum viral titer of 105.7 50% tissue culture infectious dose (TCID50)/mL in Vero cells at 5 days after inoculation. The nucleotide and amino-acid sequences of the partial segment S1 of the MReo2045 isolate were determined and compared with those of other MRV strains. The MReo2045 isolate had nucleotide sequences similar to MRV-3 and was most similar (96.1%) to the T3/Bat/Germany/342/08 strain, which was isolated in Germany in 2008. The MReo2045 isolate will be useful as an antigen for sero-epidemiological studies and developing diagnostic tools.

Identification of Void Diameters for Cast-Resin Transformers (몰드변압기의 보이드 결함 크기 판별)

  • Jeong, Gi-woo;Kim, Wook-sung
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2022.10a
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    • pp.570-573
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    • 2022
  • This paper presents the identification of void diameters for a cast-resin transformer using an artificial neural network (ANN) model. A PD signal was measured by the Rogowski coil sensor which has the planar and thin structures fabricated on a printed circuit board (PCB), and the PD electrode system was fabricated to simulate a PD defect by a void. In addition, void samples with different diameters were fabricated by injecting air in a cylindrical aluminum frame using a syringe during the epoxy curing process. To identify the diameter of void defects, PD characteristics such as the discharge magnitude, pulse count, and phase angle were extracted and back propagation algorithm (BPA) was designed using virtual instrument (VI) based on the Labview program. From the experimental results, the BPA algorithm proposed in this paper has over 90% accurate rate to identify the diameter of void defects and is expected to use reference data of maintenance and replacement of insulation for cast-resin transformers in the on-site PD measurement.

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Identification and in silico analysis of two types of serpin genes from expressed sequence tags (ESTs) of the Oriental land snail, Nesiohelix samarangae (동양달팽이 (Nesiohelix samarangae) 의 expressed sequence tags (ESTs) 로부터 분리한 2종류의 Serpin 유전자 분석)

  • Park, So Young;Jeong, Ji Eun;Hwang, Hee Ju;Wang, Tae Hun;Park, Eun Bi;Kim, Yong Min;Lee, Jun-Sang;Han, Yeon Soo;Yang, Seung-Ha;Lee, Yong Seok
    • The Korean Journal of Malacology
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    • v.30 no.2
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    • pp.155-163
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    • 2014
  • Serpins are a group of proteins involved in the regulation of serine and other type of proteases, and have been identified in many kinds of organisms from invertebrates to vertebrates. Serpins are known to regulate the proteolytic cascades of the innate immune pathways in addition to their roles in blood coagulation, angiogenesis, fibrinolysis, inflammation and tumor suppression. In this study, we have isolated two partial serpin gene fragments from expressed sequence tags (ESTs) of Nesiohelix samarangae. Dotplot analysis indicates that they are of two different types, Ns-serpin type 1 and Ns-serpin type 2. Ns-serpin type 1 has 819 bp coding region (272 amino acids), whereas Ns-serpin type 2 has 555 bp coding region (185 amino acids). Molecular phylogenetic analysis shows that the identified serpins have high similarities to their counterparts in the California see slug, Aplysia californica. Yet, the precise biological and immunological roles of these Ns-serpins remain to be further investigated using RNA interference and other molecular techniques.

Study on the Relationship among Bi-Su Type, Obesity Index, and Pattern Identification in Stroke Patients (중풍 환자에서 비수, 비만지표, 변증간 연관성에 대한 고찰)

  • Kim, So-Yeon;Lee, Jung-Sup;Kang, Byoung-Kab;Ko, Mi-Mi;Kim, Jeong-Cheol;Oh, Dal-Seok;Bang, Ok-Sun
    • The Journal of Internal Korean Medicine
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    • v.30 no.3
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    • pp.550-557
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    • 2009
  • Objectives : The purpose of this study was to investigate the possibility of Bi-Su as a pattern identification (PI) index in stroke patients. Methods : The subjects were 424 hospitalized stroke patients within 1 month from onset and diagnosed with the same PI subtypes (dampness & phlegm, qi deficiency, fire & heat, eum deficiency, and blood stasis) by agreement of two clinical experts. Bi-Su type is a kind of body shape (Bi : fat, Su : lean). Bi-Su type and degree (Bi-Su score) were decided by clinical expert. Body mass index (BMI) and waist-hip ratio (WHR) were used as an obesity index. Correlation analysis between Bi-Su score and obesity index (Spearman) and variance analysis for Bi-Su score, BMI, and WHR among PI subtypes (ANOVA) and sex were carried out. Results : While there was partial correlation between Bi-Su type and BMI($r^2$=0.634, p<0.001), the distribution of the BMI group based on the Bi-Su group showed the broadest range. The Bi-Su score in the dampness & phlegm group was higher than in the other groups (p<0.001). BMI in the dampness & phlegm groups was also higher but the BMI differences among PI subtypes was low (p=0.002). The Bi-Su score in the dampness & phlegm group was similar in both sexes, although the hand score in the eum deficiency group was the lowest, especially in males. Conclusions : Although BMI is not an objective enough tool for evaluating Bi-Su type, Bi-Su type is more appropriate than BMI as PI index. Therefore Bi-Su type could be used as one of the PI indices for dampness & phlegm or eum deficiency group in stroke patients.

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EST Analysis system for panning gene

  • Hur, Cheol-Goo;Lim, So-Hyung;Goh, Sung-Ho;Shin, Min-Su;Cho, Hwan-Gue
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2000.11a
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    • pp.21-22
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    • 2000
  • Expressed sequence tags (EFTs) are the partial segments of cDNA produced from 5 or 3 single-pass sequencing of cDNA clones, error-prone and generated in highly redundant sets. Advancement and expansion of Genomics made biologists to generate huge amount of ESTs from variety of organisms-human, microorganisms as well as plants, and the cumulated number of ESTs is over 5.3 million, As the EST data being accumulate more rapidly, it becomes bigger that the needs of the EST analysis tools for extraction of biological meaning from EST data. Among the several needs of EST analyses, the extraction of protein sequence or functional motifs from ESTs are important for the identification of their function in vivo. To accomplish that purpose the precise and accurate identification of the region where the coding sequences (CDSs) is a crucial problem to solve primarily, and it will be helpful to extract and detect of genuine CD5s and protein motifs from EST collections. Although several public tools are available for EST analysis, there is not any one to accomplish the object. Furthermore, they are not targeted to the plant ESTs but human or microorganism. Thus, to correspond the urgent needs of collaborators deals with plant ESTs and to establish the analysis system to be used as general-purpose public software we constructed the pipelined-EST analysis system by integration of public software components. The software we used are as follows - Phred/Cross-match for the quality control and vector screening, NCBI Blast for the similarity searching, ICATools for the EST clustering, Phrap for EST contig assembly, and BLOCKS/Prosite for protein motif searching. The sample data set used for the construction and verification of this system was 1,386 ESTs from human intrathymic T-cells that verified using UniGene and Nr database of NCBI. The approach for the extraction of CDSs from sample data set was carried out by comparison between sample data and protein sequences/motif database, determining matched protein sequences/motifs that agree with our defined parameters, and extracting the regions that shows similarities. In recent future, in addition to these components, it is supposed to be also integrated into our system and served that the software for the peptide mass spectrometry fingerprint analysis, one of the proteomics fields. This pipelined-EST analysis system will extend our knowledge on the plant ESTs and proteins by identification of unknown-genes.

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Species Identification and Developmental Biology of a Red Bean Pest in Ostrinia sp. (Lepidoptera: Crambidae) (Ostrinia속(나비목: 포충나방과) 팥 해충의 종 동정과 발육 특성)

  • Jung, Jin Kyo;Seo, Bo Yoon;Park, Doo-Sang;Oh, Hyun-Woo;Lee, Gwan-Seok;Park, Hae-Chul;Cho, Jum Rae
    • Korean journal of applied entomology
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    • v.51 no.4
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    • pp.469-477
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    • 2012
  • Ostrinia larvae feed the pods and stem of red bean and seriously damage the bean production from farmers. In this study we investigated biological and developmental characteristics including field collection, host feeding preference, artificial rearing diet, morphological and molecular taxonomical identification, and pheromone analysis for an Ostrinia sp. in Korea. The male adults have massive tibia in the middle legs and 3-lobed uncus in the genitalia. The partial nucleotide sequences of mitochondrial cytochrome oxidase I (COI) and II (COII) were not corresponded to those DNA sequences from other Ostrinia species reported previously in Japan and China. Host plants for this species are also different from the previous species reported. In the gas chromatography (GC) analyses, (Z)-9-tetradecenyl acetate was not detected from the pheromone gland of our species while the component as a sex pheromone was found in O. zaguliaevi and O. zealis, With taken results, we conclude this Ostrinia species in Korea is Ostrinia scapulalis or closely related species. When larvae collected in a fall were incubated in the outdoor condition, they emerged to adult between June and July in the next year. The result indicates that the winter diapause could be started in late larval stage. In addition, we developed a semi-synthetic artificial diet adopted for mass rearing of the O. scapulalis in laboratory.

Bacteriological Characteristics of Unidentified Vibrio sp., Hemolysin Producer Isolated from Brackish Water -2. Bacteriological Characteristics of Vibrio sp. E10 Similar to Vibrio mimicus- (기수에서 분리된 용혈독소를 생산하는 미분류 Vibrio sp.의 세균학적 특징 -2. Vibrio mimicus와 유사한 Vibrio sp. E10의 세균학적 특성-)

  • Kim Young Man;Yu Hong Sik;Oh Hee Kyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.6
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    • pp.545-550
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    • 2002
  • A hemolysin producing bacterial strain which belong to Vibrio species was isolated from the Kum River estuary. In the process of identification, the strain did not show characteristics of known Vibrio species; thus, the strain was designated as Vibrio sp, E10 (V. kunsan) tentatively and further identification study was carried out by comparing its bacteriological characteristics. Morphologically Vibrio sp, E10 was comma shaped rod with a polar flagellium. Clear hemolysis zones were observed with the strain against human and sheep blood agar. Hemollytic toxicity was confirmed by strong vascular Permeability and fatal toxicity against mouse was also observed. Therefore the strain was a pathogenic vibrio. Growth conditions for Vibrio sp. E10 were ranged salinity of 0$\~$$4.5\%$, pH of 6.2$\~$9.2, temperature of 14$\~$42$^{\circ}C$, respectively, 16S rDNA partial sequence of Vibrio sp, E10 showed $99\%$ homology with dozens of V. cholerae species including V, cholerae El Tor N16961 and V, snmisnfus ATCC 33653T. This strain belonged to Proteobacteria; gamma subdivision; Vibrionacea: Vibrio. But, among knorn Vibrio species no identical styains were found when using automatic bacteria identification system ($MicroLog^{TM}$system, release 4.0, Biolog Inc., USA) which evaluated the ability of metabolizing 95 kinds of carbon and nitrogen sources. Vibrio sp, E10 showed 18 and 11 different responses as compared to V. mimicus and V, cholerae, respectively.