Use of Fragments from D1/D2 Domain of 26S rRNA Gene to Select Saccharomyces cerevisiae from Palm Wine
Ogueri Nwaiwu *
Division of Food Sciences, School of Biosciences, University of Nottingham, Sutton Bonington Campus, LE12 5RD, College Road, Loughborough, Leicestershire, United Kingdom and Research Services Division, Alpha-Altis (UK) Ltd, Sir Colin Campbell Building, University of Nottingham Innovation Park, Triumph Road; Nottingham, NG7 2TU, United Kingdom
*Author to whom correspondence should be addressed.
Abstract
Aims: To determine if the 600 bp PCR amplicon of D1/D2 domain region of 26S rRNA genes in Saccharomyces cerevisiae can be used for pre-selection and identification of S. cerevisiae.
Study Design: In vitro analytical study.
Place and Duration of Study: University of Nottingham, United Kingdom; Study was carried out between March and September 2013.
Methodology: Polymerase chain reaction amplification and restriction digestion using HaeIII restriction endonuclease.
Results: The restriction profile of S. cerevisiae was different from that of Pichia kudriavzevii, Candida ethanolica and C. tropicalis. The profile was the same with known S. cerevisiae strains NCYC 1406 and S288c and yielded three fragments of approximately 306,156 and123 bp.
Conclusion: HaeIII digestion of D1/D2 domain of 26S rRNA gene of S. cerevisiae confirms that pre-selection and sequencing can be performed with one PCR reaction instead of two in palm wine yeast identification studies.
Keywords: Yeasts, restriction fragments, 26S rRNA, D1/D2 domain, palm wine, Saccharomyces cerevisiae