Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species.
Rytas Vilgalys,M Hester +1 more
TLDR
A novel approach that uses the polymerase chain reaction (PCR) for rapid simplified restriction typing and mapping of DNA from many different isolates is described, which ought to have wide applicability for clinical detection and other studies.Abstract:
Detailed restriction analyses of many samples often require substantial amounts of time and effort for DNA extraction, restriction digests, Southern blotting, and hybridization. We describe a novel approach that uses the polymerase chain reaction (PCR) for rapid simplified restriction typing and mapping of DNA from many different isolates. DNA fragments up to 2 kilobase pairs in length were efficiently amplified from crude DNA samples of several pathogenic Cryptococcus species, including C. neoformans, C. albidus, C. laurentii, and C. uniguttulatus. Digestion and electrophoresis of the PCR products by using frequent-cutting restriction enzymes produced complex restriction phenotypes (fingerprints) that were often unique for each strain or species. We used the PCR to amplify and analyze restriction pattern variation within three major portions of the ribosomal DNA (rDNA) repeats from these fungi. Detailed mapping of many restriction sites within the rDNA locus was determined by fingerprint analysis of progressively larger PCR fragments sharing a common primer site at one end. As judged by PCR fingerprints, the rDNA of 19 C. neoformans isolates showed no variation for four restriction enzymes that we surveyed. Other Cryptococcus spp. showed varying levels of restriction pattern variation within their rDNAs and were shown to be genetically distinct from C. neoformans. The PCR primers used in this study have also been successfully applied for amplification of rDNAs from other pathogenic and nonpathogenic fungi, including Candida spp., and ought to have wide applicability for clinical detection and other studies.read more
Citations
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Assessment of soil microbial community structure by use of taxon-specific quantitative PCR assays.
TL;DR: A quantitative PCR-based approach to estimating the relative abundances of major taxonomic groups of bacteria and fungi in soil provides a rapid and robust index of microbial community structure.
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Cryptococcosis in the era of AIDS--100 years after the discovery of Cryptococcus neoformans.
TL;DR: Clinical concepts are presented that relate to the distinctive features of cryptococcosis in patients with AIDS and the diagnosis, treatment, and prevention of Cryptococcus neoformans in AIDS patients.
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Fungal Molecular Systematics
TL;DR: The fungi, as thus defined, are of great importance for the following reasons: (a) They are the primary decomposers in all terrestrial ecosystems; (b) they are important symbiotic associates of vascular plants both in mutualistic and parasitic relationships.
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Taxonomy and phylogeny of Gliocladium analysed from nuclear large subunit ribosomal DNA sequences
TL;DR: The results indicate that Gliocladium is polyphyletic and that G. penicillioides, G. roseum, and Trichoderma virens, are generically distinct.
Journal ArticleDOI
Revisions to the Classification, Nomenclature, and Diversity of Eukaryotes
Sina M. Adl,David Bass,David Bass,Christopher E. Lane,Julius Lukeš,Julius Lukeš,Conrad L. Schoch,Alexey V. Smirnov,Sabine Agatha,Cédric Berney,Matthew Brown,Fabien Burki,Paco Cárdenas,Ivan Čepička,Lyudmila V. Chistyakova,Javier del Campo,Micah Dunthorn,Micah Dunthorn,Bente Edvardsen,Yana Eglit,Laure Guillou,Vladimír Hampl,Aaron A. Heiss,Mona Hoppenrath,Timothy Y. James,Anna Karnkowska,Sergey Karpov,Sergey Karpov,Eunsoo Kim,Martin Kolisko,Alexander Kudryavtsev,Daniel J. G. Lahr,Enrique Lara,Line Le Gall,Denis H. Lynn,Denis H. Lynn,David G. Mann,Ramon Massana,Edward A. D. Mitchell,Christine Morrow,Jong Soo Park,Jan Pawlowski,Martha J. Powell,Daniel J. Richter,Sonja Rueckert,Lora L. Shadwick,Satoshi Shimano,Frederick W. Spiegel,Guifré Torruella,Noha H. Youssef,Vasily V. Zlatogursky,Vasily V. Zlatogursky,Qianqian Zhang +52 more
TL;DR: It is confirmed that eukaryotes form at least two domains, the loss of monophyly in the Excavata, robust support for the Haptista and Cryptista, and suggested primer sets for DNA sequences from environmental samples that are effective for each clade are provided.
References
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TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Journal ArticleDOI
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Randall Keichi Saiki,David H. Gelfand,Susanne Stoffel,Stephen J. Scharf,Russell Higuchi,Glenn Thomas Horn,Kary B. Mullis,Henry A. Erlich +7 more
TL;DR: A thermostable DNA polymerase was used in an in vitro DNA amplification procedure, the polymerase chain reaction, which significantly improves the specificity, yield, sensitivity, and length of products that can be amplified.
Journal ArticleDOI
Mathematical model for studying genetic variation in terms of restriction endonucleases
Masatoshi Nei,Wen-Hsiung Li +1 more
TL;DR: A mathematical model for the evolutionary change of restriction sites in mitochondrial DNA is developed and a measure called "nucleotide diversity" is proposed to express the degree of polymorphism in a population at the nucleotide level.