Návrh primerů pro universální amplifikaci N-acetylglukozaminidázového genu (nag1) pro Chytridiomycety s důrazem na anaerobní Neocallimastigales

Kód výsledku v 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>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

The design of oligonucleotide primers for the universal amplification of the N-acetylglucosaminidase gene (nag1) in Chytridiomycetes with emphasis on the anaerobic Neocallimastigales

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

The design of oligonucleotide primers for the universal amplification of the N-acetylglucosaminidase gene (nag1) in Chytridiomycetes with emphasis on the anaerobic Neocallimastigales

Popis výsledku anglicky

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.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

EE - Mikrobiologie, virologie

OECD FORD obor

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Projekt

<a href="/cs/project/GA523%2F05%2F2555" target="_blank" >GA523/05/2555: Hloubková fylogeneze anaerobních hub</a><br>

Návaznosti

Z - Vyzkumny zamer (s odkazem do CEZ)

Rok uplatnění

2008

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Folia Microbiologica

ISSN

0015-5632

e-ISSN

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Svazek periodika

53

Číslo periodika v rámci svazku

3

Stát vydavatele periodika

CZ - Česká republika

Počet stran výsledku

5

Strana od-do

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Kód UT WoS článku

000257941200006

EID výsledku v databázi Scopus

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