Počet záznamů: 1
Comparative analysis of oligonucleotide primers for high-throughput screening of genes encoding adenylation domains of nonribosomal peptide synthetases in actinomycetes
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SYSNO ASEP 0451178 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Comparative analysis of oligonucleotide primers for high-throughput screening of genes encoding adenylation domains of nonribosomal peptide synthetases in actinomycetes Tvůrce(i) Bakal, Tomáš (MBU-M) RID
Goo, K.-S. (JP)
Najmanová, Lucie (MBU-M) RID
Plháčková, Kamila (MBU-M) RID
Kadlčík, Stanislav (MBU-M) RID, ORCID
Ulanová, Dana (MBU-M)Zdroj.dok. Antonie van Leeuwenhoek International Journal of General and Molecular Microbiology. - : Springer - ISSN 0003-6072
Roč. 108, č. 5 (2015), s. 1267-1274Poč.str. 8 s. Jazyk dok. eng - angličtina Země vyd. NL - Nizozemsko Klíč. slova Nonribosomal peptide synthetase ; Adenylation domain ; Actinomycetes Vědní obor RIV EE - Mikrobiologie, virologie CEP ED1.1.00/02.0109 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Institucionální podpora MBU-M - RVO:61388971 UT WOS 000362881400024 DOI 10.1007/s10482-015-0557-5 Anotace In the biosynthesis of diverse natural bioactive products the adenylation domains (ADs) of nonribosomal peptide synthetases select specific precursors from the cellular pool and activate them for further incorporation into the scaffold of the final compound. Therefore, the drug discovery programs employing PCR-based screening studies of microbial collections or metagenomic libraries often use AD-coding genes as markers of relevant biosynthetic gene clusters. However, due to significant sequence diversity of ADs, the conventional approach using only one primer pair in a single screening experiment could be insufficient for maximal coverage of AD abundance. In this study, the widely used primer pair A3F/A7R was compared with the newly designed aa194F/aa413R one by 454 pyrosequencing of two sets of actinomycete strains from highly dissimilar environments: subseafloor sediments and forest soil. Individually, none of the primer pairs was able to cover the overall diversity of ADs. However, due to slightly shifted specificity of the primer pairs, the total number and diversity of identified ADs were noticeably extended when both primer pairs were used in a single assay. Additionally, the efficiency of AD detection by different primer combinations was confirmed on the model of Salinispora tropica genomic DNA of known sequence Pracoviště Mikrobiologický ústav Kontakt Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Rok sběru 2016
Počet záznamů: 1