Identification of an ARS element and development of a high efficiency transformation system for Pichia guilliermondii

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 f...

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Bibliographic Details
Published in:Current genetics 1999-10, Vol.36 (4), p.215-221
Main Authors: Boretsky, Y, Voronovsky, A, Liuta-Tehlivets, O, Hasslacher, M, Kohlwein, S.D, Shavlovsky, G.M
Format: Article
Language:English
Subjects:
3' Untranslated Regions
Amino Acid Sequence
amino acid sequences
autonomous replication
autonomously replicating sequence
Base Sequence
Blotting, Southern
Cloning, Molecular
DNA replication
genbank/z49093
genes
Genes, Fungal
genetic transformation
genetic vectors
GTP Cyclohydrolase - genetics
gtp cyclohydrolase ii
hydrolases
mitosis
Molecular Sequence Data
molecular seuqence data
nucleotide sequences
open reading frames
Pichia
Pichia - enzymology
Pichia - genetics
Pichia guilliermondii
plasmid vectors
Plasmids
Replication Origin
rib 1 gene
RIB1 gene
shuttle vectors
Transformation, Genetic
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Summary: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 10(4)-10(5) transformants/microg 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.
ISSN:0172-8083
1432-0983
DOI:10.1007/s002940050493