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Prikladnaia biokhimiia i mikrobiologiia

Cloning and bioinformatic analysis of an acidophilic beta-mannanase gene, Anman5A, from Aspergillus niger LW-1.


PMID 23101390

Abstract

Using 3' and 5' rapid amplification of cDNA ends (RACE) techniques, the full-length cDNA sequence of the AnmanSA, a gene that encodes an acidophilic beta-mannanase of Aspergillus niger LW-1 (abbreviated to AnMan5A), was identified from the total RNA. The cDNA sequence was 1417 bp in length, harboring 5'- and 3'-untranslated regions, as well as an open reading frame (ORF) which encodes a 21-aa signal peptide, a 17-aa propeptide and a 345-aa mature peptide. Based on the topology of the phylogenetic tree of beta3-mannanases from glycoside hydrolase (GH) family 5, the AnMan5A belongs to the subfamily 7 of the GH family 5. Its 3D structure was modeled by the bitemplate-based method using both MODELLER 9.9 and SALIGN programs, based on the known beta-mannanase crystal structures of Trichoderma reesei (1QNO) and Lycopersicon esculentum (1RH9) from the GH family 5. In addition, the complete DNA sequence of the Anman5A was amplified from the genomic DNA using the pUCm-T vector-mediated PCR and conventional PCR methods. The DNA sequence was 1825 bp in length, containing a 5'-flanking regulatory region, 2 introns and 3 exons when compared with the full-length cDNA.

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