1.Identification of Some Phellinus spp..
Mycobiology 2001;29(4):190-193
Four strains of Phellinus spp. was identified based on internal transcribed spacer (ITS) region of rDNA sequence analysis and morphological characteristics. Basidiocarps of all strains were effused-reflexed and hymenial surface was poroid. Hyphal system was dimitic and basidiospore was globose to ellipsoid. The amplification of ITS regions produced a DNA fragment of 500 to 780 bp in all strains examined. The determined sequences were analyzed for the reconstruction of phylogenetic tree. From these results, Phellinus sp. KM-1, KM-2, and KM-4 was identified as P. hartigii, P. baumii, and P. linteus, respectively.
DNA
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DNA, Ribosomal
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Fruiting Bodies, Fungal
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Sequence Analysis
2.Characterization of Sporothrix schenckii by random amplification of polymorphic DNA assay.
Xiaoming LIU ; Cuihong LIAN ; Liji JIN ; Lijia AN ; Guoling YANG ; Xiran LIN
Chinese Medical Journal 2003;116(2):239-242
OBJECTIVESTo investigate the DNA polymorphism of Sporothrix schenckii (S. schenckii) and to find the relationship between DNA patterns and geographic areas and clinical manifestations.
METHODThe total DNA was extracted with hexadecyltrimethyl-ammonium bromide. Random Amplification of Polymorphic DNA (RAPD) assay was used to study DNA typing of 24 strains of S. schenckii collected from different areas and isolated from different clinical types.
RESULTSOf seven random primers used, three primers (OPAA11, OPD18 and OPB07) gave good reactions, the sequences of which were 5'-ACCCGACCTG-3', 5'-GAGAGCCAAC-3', 5'-GGTGAC~GCAG-3' respectively. The RAPD patterns of the 24 isolates were not completely identical, showing certain degrees of hereditary variability. Different isolates showed a common conserved DNA band with the same primer. Different clinical types showed different genotypes.
CONCLUSIONRAPD analysis is useful in DNA typing of S. schenckii, the DNA band type of which is related to geographic origin and Clinical manifestation.
DNA, Fungal ; analysis ; Humans ; Random Amplified Polymorphic DNA Technique ; Sporothrix ; genetics
3.Identification of common species of dermatophytes by PCR-RFLP.
Ganlin, HE ; Jiawen, LI ; Juan, DING ; Zhijan, TAN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2005;25(4):458-60
To establish a simple, sensitive and effective technique for the identification of six common dermatophytes, polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) targeting Topoisomerase II gene were used. The DNA of 6 common dermatophytes was amplified by primer dPsD1 and then primers dPsD2. The products generated by dPsD2 were digested with restriction enzyme Hinc II and Hinf I separately. A DNA fragment of about 3390 bp was amplified by using primer dPsD1 from the genomic DNA of each dermatophyte species. The product of dPsD2 was 2380 bp and the restriction profiles of Hinc II and Hinf I were between 58-1670 bp. By using PCR-RFLP, all of the 6 dermatophytoses were diagnosed to species level and no obvious difference identification between Hinc II and Hinf I. It is concluded that the PCR-RFLP identification of dermatophytes by Hinc II or Hinf I is efficient and rapid in clinical practice.
Arthrodermataceae/*classification
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Arthrodermataceae/genetics
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Arthrodermataceae/*isolation & purification
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DNA Topoisomerases, Type II/genetics
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DNA, Fungal/analysis
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DNA, Fungal/genetics
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Dermatomycoses/*microbiology
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Polymerase Chain Reaction
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Polymorphism, Restriction Fragment Length
4.Sequence Analyses of PCR Amplified Partial SSU of Ribosomal DNA for Identifying Arbuscular Mycorrhizal Fungi in Plant Roots.
Moon Sung TAE ; Ahn Heum EOM ; Sang Sun LEE
Mycobiology 2002;30(1):13-17
The genomic DNAs were extracted from roots of Glycine max and Sorghum bicolor, and compared with those from spores of two arbuscular mycorrhizal (AM) fungi, Glomus mosseae and Scutellospora heterogama. The partial small subunit (SSU) of ribosomal RNA genes were synthesized and amplified by polymerase chain reaction with the fungal specific primers, AM1 and NS31. By the recent molecular techniques, the presence of another AM fungal DNA were confirmed in the roots of two plants, and three sequences of rDNA fragments amplified were identified to be close to those of G. caledonium, G. fasiculatum and G. proliferum. The two AM fungi, both, were found to colonize at the cortical layers of plant roots collected in the fields, together.
Colon
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DNA
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DNA, Fungal
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DNA, Ribosomal*
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Fungi*
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Genes, rRNA
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Plant Roots*
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Plants*
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Polymerase Chain Reaction*
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Sequence Analysis*
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Sorghum
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Soybeans
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Spores
5.Representative appressorium stage cDNA library of Magnaporthe grisea.
Jian-ping LU ; Tong-bao LIU ; Xiao-yun YU ; Fu-cheng LIN
Journal of Zhejiang University. Science. B 2005;6(2):132-136
A mature appressorium cDNA library of rice blast fungus, Magnaporthe grisea, was constructed in a lambdaTriplEx2 vector by SMART cDNA library containing 2.37x10(6) independent clones about 100% of which harbor foreign cDNA inserts with average size of 660 bp. Of 9 randomly selected clones, 2 expressed sequence tags (ESTs) sequences did not have homologous EST sequences of M. grisea in GenBank. The appressorium cDNA library is suitable for gene expression analysis and function analysis of the late stages of appressorium formation and the early stages of penetration of M. grisea.
Cloning, Molecular
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methods
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DNA, Fungal
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genetics
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Gene Expression Profiling
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methods
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Gene Expression Regulation, Fungal
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Gene Library
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Magnaporthe
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genetics
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Sequence Analysis, DNA
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methods
6.DNA and RNA random amplification polymorphism in 5-flurocytosine-resistant strains of Candida albicans from revul-vaginal candidasis.
Journal of Zhejiang University. Medical sciences 2005;34(1):69-72
OBJECTIVETo investigate the DNA genome and RNA expression in 5-flurocytosine-resistant strains of Candida albicans from vaginal candidasis.
METHODSSixteen strains of Candida albicans were selected from clinically diagnosed revul-vaginal candidasis. Eight 5-flurocytosine-sensitive isolates and 8 resistant isolates were examined by France Media FUNGUS sensitive test. DNA genome was detected with random amplification polymorph DNA. RNA expression was detected with random amplification polymorph RNA method.
RESULTSThere were no distinct differences between 5-flurocytosine-sensitive and resistant Candida albicans in DNA genome, while RNA expression showed significant differences between 5-flurocytosine-resistant and sensitive strains.
CONCLUSIONClinical 5-flurocytosine-resistant strains of Candida albicans from revul-vaginal candidasis may be related to phenotype changes.
Adolescent ; Adult ; Antifungal Agents ; pharmacology ; Candida albicans ; drug effects ; genetics ; Candidiasis, Vulvovaginal ; microbiology ; DNA, Fungal ; analysis ; Drug Resistance, Fungal ; Female ; Flucytosine ; pharmacology ; Humans ; Middle Aged ; RNA, Fungal ; analysis ; Random Amplified Polymorphic DNA Technique
7.AFLP analysis for genetic diversity of Ganoderma.
Lin-yong ZHENG ; Ding-hong JIA ; Xia LUO ; Zhi-rong YANG
China Journal of Chinese Materia Medica 2007;32(17):1733-1736
OBJECTIVETo investigate the genetic diversity of Ganoderma cultivars provided for the genuineness study, germ-plasm resource identification, genetic relationship study, breeding, introduction and cultivante of Ganoderma strains.
METHODWith the software of NTSYSpc 2. 1, 24 materials, of G. lucidum and G. sinense, were studied using AFLP to construct the dendrogram.
RESULTThere were 177 polymorphic bands with 14 primer combinations. And all materials could be identified with AFLP.
CONCLUSIONThere actually existed much genetic diversity at the molecular level among the germplasm resources of Ganoderma strains, and all the strains were clustered into G. lucidum group and G. sinense group at the similarity coefficient 0. 676.
Amplified Fragment Length Polymorphism Analysis ; China ; Cluster Analysis ; DNA Primers ; DNA, Fungal ; analysis ; genetics ; Ganoderma ; classification ; genetics ; Genetic Variation ; Phylogeny ; Reishi ; genetics
8.Physical interactions and mutational analysis of MoYpt7 in Magnaporthe oryzae.
Lu-Yao HUANG ; Min WU ; Xiao-Yun YU ; Lin LI ; Fu-Cheng LIN ; Xiao-Hong LIU
Journal of Zhejiang University. Science. B 2018;19(1):79-84
In this study, we analyzed the physical interactions of the dominant negative isoform of MoYpt7. Our results show that MoYpt7 interacts with MoGdi1. The dominant negative isoform of MoYpt7 (dominant negative isoform, N125I) is essential for colony morphology, conidiation, and pathogenicity in the rice blast fungus. These results further demonstrate the biological functions of MoYpt7 in Magnaporthe oryzae.
DNA Mutational Analysis
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Fungal Proteins/metabolism*
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Gene Expression Regulation, Fungal
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Genes, Fungal
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Green Fluorescent Proteins/metabolism*
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Magnaporthe/genetics*
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Microscopy, Fluorescence
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Mutation
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Oryza/microbiology*
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Phenotype
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Plant Diseases/microbiology*
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Protein Isoforms
9.Molecular phylogenetic analysis of Paecilomyces hepiali and Cordyceps sinensis.
Jin-Ling YANG ; Wei XIAO ; Hui-Xia HE ; Hui-Xin ZHU ; Shu-Fang WANG ; Ke-Di CHENG ; Ping ZHU
Acta Pharmaceutica Sinica 2008;43(4):421-426
Phylogenetic relationship between Paecilomyces hepiali and Cordyceps sinensis was studied by analyzing the sequence of rDNA-ITS. The samples of C. sinensis were collected from Hualong County in Qinghai Province and Kangding County in Sichuan Province in May and June, respectively. The rDNA-ITS fragments were obtained by PCR amplification with the template genomic DNA of the fresh stroma or caterpillar body of the collected samples and the cultured mycelium of P. hepiali, with the universal fungal primers ITS1/ITS4. The amplified fragments were cloned into pMD18-T Vector and sequenced. Phylogenetic analysis was performed with these sequences and those from GenBank. The result showed that all of the 46 clones randomly chosen from the amplification of C. sinensis shared identical or almost identical rDNA-ITS regions and had over 99% identity with some rDNA-ITS sequences of Hirsutella sinensis and C. sinensis registered in GenBank, but all of them had only about 72% identity with that of P. hepiali. Two pairs of specific primers were designed based on the rDNA-ITS sequence of P. hepiali, then PCR and Nest-PCR were performed with the template genomic DNA of the stroma or caterpillar body of C. sinensis samples mentioned above. The apparent bands amplified by Nest-PCR were obtained from all of the samples, and the sequences showed 100% identity with the rDNA-ITS sequence of P. hepiali. In addition, another pair of specific primers were designed based on the rDNA-ITS sequence registered in GenBank as the marker of C. sinensis (accession no. AB067740) but the latter only shared 87.3% identity with that of H. sinensis (accession no. AJ309353). This pair of primers was used to amplify the C. sinensis samples by PCR, and the amplified sequence showed 100% identity with that of AB067740. The result indicated that H. sinensis is the main body of C. sinensis, while some other endoparasitic fungi such as P. hepiali commonly exist in the natural C. sinensis.
Base Sequence
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Cordyceps
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classification
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genetics
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DNA, Fungal
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genetics
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DNA, Ribosomal Spacer
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genetics
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Molecular Sequence Data
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Paecilomyces
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classification
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genetics
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Phylogeny
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Polymerase Chain Reaction
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methods
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Sequence Alignment
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Sequence Analysis, DNA
10.Scaling behavior of nucleotide cluster in DNA sequences.
Jun CHENG ; Zi-shuang TONG ; Lin-xi ZHANG
Journal of Zhejiang University. Science. B 2007;8(5):359-364
In this paper we study the scaling behavior of nucleotide cluster in 11 chromosomes of Encephalitozoon cuniculi Genome. The statistical distribution of nucleotide clusters for 11 chromosomes is characterized by the scaling behavior of P(S) proportional, variant e(-alphaS), where S represents nucleotide cluster size. The cluster-size distribution P(S(1)+S(2)) with the total size of sequential C-G cluster and A-T cluster S(1)+S(2) were also studied. P(S(1)+S(2)) follows exponential decay. There does not exist the case of large C-G cluster following large A-T cluster or large A-T cluster following large C-G cluster. We also discuss the relatively random walk length function L(n) and the local compositional complexity of nucleotide sequences based on a new model. These investigations may provide some insight into nucleotide cluster of DNA sequence.
Base Sequence
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Computer Simulation
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DNA, Fungal
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genetics
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Encephalitozoon cuniculi
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genetics
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Evolution, Molecular
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Models, Genetic
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Molecular Sequence Data
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Multigene Family
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genetics
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Nucleotides
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genetics
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Sequence Analysis, DNA
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methods