The design of oligonucleotide primers for the universal amplification of the N-acetylglucosaminidase gene (nag1) in Chytridiomycetes with emphasis on the anaerobic Neocallimastigales
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985904%3A_____%2F08%3A00324753" target="_blank" >RIV/67985904:_____/08:00324753 - isvavai.cz</a>
Result on the web
—
DOI - Digital Object Identifier
—
Alternative languages
Result language
angličtina
Original language name
The design of oligonucleotide primers for the universal amplification of the N-acetylglucosaminidase gene (nag1) in Chytridiomycetes with emphasis on the anaerobic Neocallimastigales
Original language description
The common feature of all chytridiomycetous fungi, aerobic as well as anaerobic, is abundance of chitin in their cell wall. Genes encoding chitinases were widely used as phylogenetic markers in ascomycetes, but their utility was not tested for Chytridiomycetes. In this study we chose the gene encoding an enzyme involved in chitin degradation and energy metabolism, the b-(1, 4)-N-acetyl-glucosaminidase (Nag1). Primer pair Nag-fwd and Nag-rev was used to create PCR product from five strains of anaerobic and seven strains of aerobic chytrids. Blast search of sequenced amplicons however proved that these primers are specific only for fungus Emericella nidulans. Amino acid alignement of Nag1 proteins of fungal, protozoal and bacterial origin available in GenBank database was therefore performed in this work. Five amino acid regions were found to be conserved enough to serve as suitable domain for the design of set of primers for the universal amplification of the Nag1 gene in fungi.
Czech name
Návrh primerů pro universální amplifikaci N-acetylglukozaminidázového genu (nag1) pro Chytridiomycety s důrazem na anaerobní Neocallimastigales
Czech description
Přítomnost chitinu v buněčných stěnách je charakteristická pro mnoho hub včetně Chytridimycet. Tyto geny proto mají velký potenciál ve fylogenetickém využití, dosud však byly aplikovány hlavně u Askomycet, ale u Chytridiomycet nebyly použity. Aplikovalijsme proto primery Nag-forward a Nag-reverse na 5 anaerobních a 7 anaerobních chytridií. Experimenty prokázaly, že tyto primery jsou specifické pouze pro Emericella nudilans. Proto na základě konservativních sekvencí fungálních, protozálních a bakteriálních proteinů pro Nag1 z Genové Banky byl navržen set primerů pro universální amplifikaci genu nag1 u hub třídy Neocallimastigales.
Classification
Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
EE - Microbiology, virology
OECD FORD branch
—
Result continuities
Project
<a href="/en/project/GA523%2F05%2F2555" target="_blank" >GA523/05/2555: Deep phylogeny of anaerobic fungi</a><br>
Continuities
Z - Vyzkumny zamer (s odkazem do CEZ)
Others
Publication year
2008
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Folia Microbiologica
ISSN
0015-5632
e-ISSN
—
Volume of the periodical
53
Issue of the periodical within the volume
3
Country of publishing house
CZ - CZECH REPUBLIC
Number of pages
5
Pages from-to
—
UT code for WoS article
000257941200006
EID of the result in the Scopus database
—