• Title/Summary/Keyword: Species similarity

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Cloning and Expression in Pichia pastoris of a New Cytochrome P450 Gene from a Dandruff-causing Malassezia globosa

  • Lee, Eun-Chang;Ohk, Seul-Ong;Suh, Bo-Young;Park, Na-Hee;Kim, Beom-Joon;Kim, Dong-Hak;Chun, Young-Jin
    • Toxicological Research
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    • v.26 no.1
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    • pp.47-52
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    • 2010
  • The Malassezia fungi are responsible for various human skin disorders including dandruff and seborrheic dermatitis. Of the Malassezia fungi, Malassezia globosa (M. globosa) is one of the most common in human scalp. The completed genome sequence of M. globosa contains four putative cytochrome P450 genes. To determine the roles of Malassezia P450 enzymes in the biosynthesis of ergosterol, we isolated MGL3996 gene from M. globosa chromosomal DNA by PCR. The MGL3996 gene encodes an enzyme of 616 amino acids, which shows strong similarity with known CYP52s of other species. MGL3996 gene was cloned and expressed in Pichia pastoris (P. pastoris) heterologous yeast expression system. Using the yeast microsomes expressing MGL3996 protein, a typical P450 CO-difference spectrum was shown with absorption maximum at 448 nm. SDS-PAGE analysis revealed a protein band of apparent molecular weight 69 kDa and Western blot with anti-histidine tag antibody showed that MGL3996 was successfully expressed in P. pastoris. Cloning and expression of a new P450 gene is an important step to study the P450 monooxygenase system of M. globosa and to understand the role of P450 enzymes in pathophysiology of dandruff.

Development of Molecular Markers for Alternative Oxidase Synthesis Genes in Brassica rapa L. (배추 alternative oxidase 합성 유전자와 연관된 분자마커 개발)

  • Jeong, Ye-Sol;Chung, Sang-Min
    • Journal of Life Science
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    • v.20 no.2
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    • pp.208-212
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    • 2010
  • The low and high temperature stress might affect the yield and quality of many crop species. Alternative oxidase (AOX) gene is known as factors related to stress resistance in plants. In order to develop molecular markers related to stress resistance in Chinese cabbage, fifteen ESTs sharing sequence similarity to arabidopsis AOX genes were found using Brassica rapa EST database from NCBI. The polymorphic DNA sequences using the ESTs were then screened between Chinese cabbage, 'Chiifu' and 'Kenshin'. We found four ESTs that have either insertion or deletion between the two cultivars. These polymorphic sites were then targeted for development of the four PCR based molecular markers. These molecular markers developed in this study could be useful for a test of their relationship with abiotic stress resistance in Chinese cabbage.

A Study on the Taxonomic Status for Nitrogen-Fixing, Methanol Utilizing Oligotrophic Bacteria (저영양세균중(低營養細菌中) 질소고정균(窒素固定菌) 및 메타놀이용균(利用菌)의 분류학적위치(分類學的位置)에 관(關)하여)

  • Shin, Gawan Chull;Whang, Kyung Sook;Hattori, Tsutomu
    • Korean Journal of Agricultural Science
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    • v.16 no.2
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    • pp.163-168
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    • 1989
  • Fourty-four isolates showed acetylene-reducing(nitrogenase)activity under the atmosphere of 89% Ar, 10% $C_2H_2$ and 1 % $O_{2{\cdot}}$, these nitrogen-fixing isolates characterized chemotaxonomically and their taxonomic status was disscussed; twenty-three isolates corresponded to Azospirillum. They were curved/spiral rods, gram negative, motile by a polar flagellum, and also utilized glucose in nitrogen free medild by a polar flagellum, and also utilized glucose in nitrogen free medium. but the cellular fatty acid composition and quinone system of these isolates showed quite different characteristics with reference strains. Therefore, the taxonomic status of this nitrogen-fixing bacteria is disscussed and a new species Azospirillum. Sixty forur isolates utilized C-l compounds such as methanol and formic acid. phenotypic and chemotaxonomic characteristics of methanol utilizing isolates were investigated and their taxonomic status was discussed; Twenty-one isolates corresponded to Hyphomicrobium and for the other regular rods and irregular rods utilizing isolates showed different cellular fatty acid composition. These isolates were grouped into 8 cluster analysis and similarity values based on correlation coefficients. Among these 8 clusters, two corresponded Pseudomonas and for the other were not decided.

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User Satisfaction Analysis on Similarity-based Inference Insect Search Method in u-Learning Insect Observation using Smart Phone (스마트폰을 이용한 유러닝 곤충관찰학습에 있어서 유사곤충 추론검색기법의 사용자 만족도 분석)

  • Jun, Eung Sup
    • Journal of the Korea Society of Computer and Information
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    • v.19 no.1
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    • pp.203-213
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    • 2014
  • In this study, we proposed a new model with ISOIA (Insect Search by Observation based on Insect Appearance) method based on observation by insect appearance to improve user satisfaction, and compared it with the ISBC and ISOBC methods. In order to test these three insect search systems with AHP method, we derived three evaluation criteria for user satisfaction and three sub-evaluation criteria by evaluation criterion. In the ecological environment, non-experts need insect search systems to identify insect species and to get u-Learning contents related to the insects. To assist the public the non-experts, ISBC (Insect Search by Biological Classification) method based on biological classification to search insects and ISOBC (Insect Search by Observation based on Biological Classification) method based on the inference that identifies the observed insect through observation according to biological classification have been provided. In the test results, we found the order of priorities was ISOIA, ISOBC, and ISBC. It shows that the ISOIA system proposed in this study is superior in usage and quality compared with the previous insect search systems.

Comparative studies of the five edible mountain vegetables by DNA marker fingerprinting (DNA marker 지문법에 의한 취나물 5종 (청옥취 , 개미취 , 참취 , 수리취 , 곰취)의 비교연구)

  • 유기억
    • Korean Journal of Plant Resources
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    • v.9 no.3
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    • pp.305-310
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    • 1996
  • Five edible mountain vegetables(Saussurea sp. Aster tataricus, A. scaber. Synurus deltoides, Ligularia fischeri) were investigated on the basis of amplified DNA polymorphisms resulted from PCR (polymerase chain reaction) analysis. The sampled plants consisted of 38 individuals in 5 taxa. Only 10 primers out of 62 primers (60 random [10-mer] primers, two 15-mer [M13 core sequence, and (GGAT) sequence]) tested gave rise to polymorphisms in all of the tested plants, producing 176 DAN fragments amplified. Intraspecific polymorphisms found in each taxa showed intraspecies constancy (31.1-61.1%) in the banding patterns of individual plants: Saussurea sp. 31.1%, 15 bands, Aster tataricus, 40.9%, 18 bands, A. scaber. 38.5%, 15 bands. Synurus deltoides, 34.7%, 17 bands, and Ligularia fischeri, 61.1%, 22 brands, respectively. All five species were well classified from each other at the 0.93 level of similarity index value. Intraspecific and interspecific variations were appeared at the levels ranging from 0.62 to 0.99. Based on these results, our PCR analyses support the previous data derived from external morphology of the 5 edible mountain vegetables, but very low levels o intraspecific variations were detected in all of these taxa.

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Genetic diversity of conserved potato germplasm using microsatellite markers

  • Lee, Gi-An;Cho, Kwang-Soo;Shin, Myoung-Jae;Lee, Jung-Ro;Cho, Yang-Hee;Ma, Kyung-Ho
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.85-85
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    • 2017
  • Potato is important carbohydrate source over the world in that revealing high productivity per the unit area, and their cultivation area is estimated to be increased to cope with a scarcity of food according to the population increase. Major cultivated species of potato is Solanum tuberosum (2n = 4x = 48) and regarded as being originated in Andes region of South America. The diverse potato genetic resources has been collected and perserved in Highland Agricultural Research Institute (NICS, RDA), and the genetic materials as DNA stock is conserved in National Agrobiodiversity Center(NAS, RDA). The understanding of genetic constitution of conserved diversity is the basis for the germplam management and further utilization. In this study, we analyzed the genetic diversity of potato germplasm(479 accessions) using 24 microsatellite markers which have been internationally used for fingerprinting of potato accession. The allele number and polymorphic information content (PIC) of total accessions per locus was ranged from 2 to 18 (mean = 8.2) and from 0.214 to 0.771 (mean = 0.595), respectively. Especially, the accession originated from Korea revealed average allele number of 6.0 (2 - 11) and average PIC value of 0.58 (0.193 - 0.763). Three groups were deduced by phylogenic analysis (Group-1, -2, -3); Korean accessions showed close genetic similarity to Japanese and USA accessions, and Korean landraces were mainly included in Group-3. We try to elaborate the genetic diversity analysis of conserved potato germplasm by acquiring more genotypes using applicable molecular markers.

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Molecular Characterization of Epoxide Hydrolase from Aspergillus niger LK using Phylogenetic Analysis (진화적 유연관계 분석을 통한 Aspergillus niger LK의 Epoxide Hydrolase의 특성분석)

  • 김희숙;이은열;이수정;이지원
    • KSBB Journal
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    • v.19 no.1
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    • pp.42-49
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    • 2004
  • A gene coding for epoxide hydrolase (EH) of Aspergillus niger LK, a fungus possessing the enantioselective hydrolysis activity for racemic epoxides, was characterized by phylogenetic analysis. The deduced protein of A. niger LK epoxide hydrolase shares significant sequence similarity with several bacterial EHs and mammalian microsomal EHs (mEH) and belongs to the a/${\beta}$ hydrolase fold family. EH from A. niger LK had 90.6% identity with 3D crystal structure of lqo7 in Protein Data Bank. Sequence comparison with other source EHs suggested that Asp$\^$l92/, Asp$\^$374/ and His$\^$374/ constituted the catalytic triad. Based on the multiple sequence comparison of the functional and structural domain sequence, the phylogenetic tree between relevant epoxide hydrolases from various species were reconstructed by using Neighbor-Joining method. Genetic distances were so far as 1.841-2.682 but characteristic oxyanion hole and catalytic triad were highly conserved, which means they have diverged from a common ancestor.

Screening of Specific Genes Expressed in the Swine Tissues and Development of a Functional cDNA Chip

  • Kim, Chul Wook;Chang, Kyu Tae;Hong, Yeon Hee;Kwon, Eun Jung;Jung, Won Yong;Cho, Kwang Keun;Chung, Ki Hwa;Kim, Byeong Woo;Lee, Jung Gyu;Yeo, Jung-Sou;Kang, Yang Su;Joo, Young Kuk
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.7
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    • pp.933-941
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    • 2005
  • To develop a functional cDNA chip, specific genes expressed in the tissues of swine Kagoshima Berkshire were screened. A total of 4,434 ESTs were obtained by constructing a cDNA library from total RNA isolated from the muscle and fat tissues, affirming their functions by investigating similarity of nucleotide sequences with the database at the NCBI. Among them, 1,230 ESTs were confirmed as novel genes, which, to date, have not been identified. Attaching the genes to a cDNA microarray slide revealed expression patterns of genes in muscle and fat according to the growth stages of swine. As specific genes expressed in the muscle tissues of swine with body weight of 30 kg, 60 genes including actin, myosin, tropomysin, transfer RNA-trp synthetase, Kel-like protein 23, KIAA0182 and COI, Foocen-m, etc were obtained. In addition, 18 novel genes were obtained. As specific genes expressed in fat tissues of swine with body weight of 30 kg, 47 genes including annexin II, Collagen, Fibronectin, Pleckstrin homology domain, serine protease, etc were obtained. 21 novel genes were also obtained. The genes specifically expressed in the muscle and fat tissues of swine affect contraction and relaxation of the muscle and the fat. However, studies on the expression mechanisms of the genes are insufficient. To reveal species of structural genes in swine muscle and fat tissue, interrelation studies in expression and function of genes by using the cDNA chip should be conducted.

Discrimination and Genetic Relationship of Adenophorae triphylla(Thunb) A.DC. var. japonica Hara and Codonopsis lanceolata Trauty using RAPD analysis (RAPD분석에 의한 잔대와 더덕의 유연관계 비교 및 감별)

  • Lee, Mi-Young;Mo, Suk-Yeon;Kim, Du-Whan;Oh, Seong-Eun;Ko, Byoung-Seob
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.3
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    • pp.205-210
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    • 2001
  • Dried parts of the two species are difficult to distinguish morphologically, thus Codonopsis radix has been sold instead of Adenophorae radix in herbal medicine market. Therefore, this study was conducted to develop the genetic marker through the examination of the phylogenetic relationships between two Adenophora triphylla(Thunb.) A. DC. var. japonica Hara, two Adenophora radiatifolia Nakai, five Codonopsis lanceolata(Sieb. et Zucc)Trautv. using RAPD analysis. Fifty decarmer oligonucleotide primers were screened for the RAPD analysis, and four primers generated distinct RAPD markers specific to Adenophorae radix and Codonopsis radix. Based on the RAPD patterns, the genetic relationships between three herbal medicine were analyzed by UPGMA method. As a result, Adenophorae radix and Codonopsis radix were classified into two major subgroups on the basis of the genetic similarity coefficient. The specific RAPD patterns generated by the selected primers were reproducible from dried materials. Furthermore, the specific RAPD patterns were produced from the mixture of dried roots of A. triphylla and C. lanceolata. These results prone the usefulness of the RAPD analysis for the discrimination of pure materials from the mixtures of A. triphylla and C. lanceolata.

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Molecular Divergences of 16S rRNA and rpoB Gene in Marine Isolates of the Order Oscillatoriales (Cyanobacteria) (남조세균 흔들말목(Cyanobacteria, Oscillatoriales) 해양 균주의 16S rRNA와 rpoB 유전자 변이)

  • Cheon, Ju-Yong;Lee, Min-Ah;Ki, Jang-Seu
    • Korean Journal of Microbiology
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    • v.48 no.4
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    • pp.319-324
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    • 2012
  • In this study, we investigated molecular divergences and phylogenetic characteristics of the 16S ribosomal RNA (rRNA) and RNA polymerase beta subunit (rpoB) gene sequences from the order Oscillatoriales (Cyanobacteria). The rpoB of Oscillatoriales showed higher genetic divergence when compared with those of 16S rRNA (p-distance: rpoB=0.270, 16S=0.109), and these differences were statistically significant (Student t-test, p<0.001). Phylogenetic trees of 16S rRNA and rpoB were generally compatible; however, rpoB tree clearly separated the compared Oscillatoriales taxa, with higher phylogenetic resolution. In addition, parsimony analyses showed that rpoB gene evolved 2.40-fold faster than 16S rRNA. These results suggest that the rpoB is a useful gene for the molecular phylogenetics and species discrimination in the order Oscillatoriales.