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Identification of an ARS element and development of a high efficiency transformation system for Pichia guilliermondii |
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| Authors: | Yuriy Boretsky Andriy Voronovsky Oksana Liuta-Tehlivets Meinhard Hasslacher Sepp D. Kohlwein Georgiy M. Shavlovsky |
| Affiliation: | (1) Division of Regulatory Cell Systems, A.V. Palladin Institute of Biochemistry, Dragomanov St., 14-16, 290005 Lviv, Ukraine, UA;(2) SFB Biomembrane Research Center, Department of Biochemistry, Technical University Graz, Petersgasse 12, A-8010 Graz, Austria e-mail: liauta@kfunigraz.ac.at Tel.: +43-316 873 6458 Fax: +43-316 873 6952, AT |
| Abstract: | An Autonomously Replicating Sequence element adjacent to the RIB1 gene encoding GTP cyclohydrolase II of the yeast Pichia guilliermondii was identified by transformation experiments. Detailed sequence analysis unveiled two potential ARS elements located 5′ and 3′ of the RIB1 open reading frame. The chromosomal fragment containing the ARS-like sequence 3′ to the RIB1 structural gene, called PgARS, conferred high transformation frequencies of 104–105 transformants/μg of DNA to a pUC19-derived plasmid in P. guilliermondii. The PgARS element also conferred autonomous replication to hybrid plasmids in this host. Based on this element a series of Escherichia coli shuttle vectors for efficient transformation of the flavinogenic yeast P. guilliermondii was developed. Received: 22 September 1998 / 1 June 1999 |
| Keywords: | Autonomously replicating sequence Flavinogenic yeast GTP cyclohydrolase II Transformation system |
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