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Boundaries in metagenomic screenings using lacZα-based vectors Genet. Mol. Biol.
Alves,Luana de Fátima; Borelli,Tiago Cabral; Westmann,Cauã Antunes; Silva-Rocha,Rafael; Guazzaroni,María-Eugenia.
Abstract Metagenomics approaches have been of high relevance for providing enzymes used in diverse industrial applications. In the current study, we have focused on the prospection of protease and glycosyl hydrolase activities from a soil sample by using the lacZα -based plasmid pSEVA232. For this, we used a functional screen based on skimmed milk agar and a pH indicator dye for detection of both enzymes, as previously reported in literature. Although we effectively identified positive clones in the screenings, subsequent experiments revealed that this phenotype was not because of the hydrolytic activity encoded in the metagenomic fragments, but rather due to the insertion of small metagenomic DNA fragments in frame within the coding region of the lacZ...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Functional metagenomics; Protease; Glycosyl hydrolase; False positive clones.
Ano: 2020 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572020000100803
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