Biopolymers and cell. 2000; 16 (1): 46 - 52

Identification of an autonomously replicating sequence of the flavinogenic yeast Pichia guilliermondii

Andriy Voronovsky, Yuriy Boretsky

An autonomously replicating sequence adjacent to the RfBl 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 RIB I open reading frame. The chromosomal fragment containing the ARS-like sequence 3' to the RIBJ structural gene, called PgARS, conferred autonomous replication to hybrid plasmids and high transformation frequencies (10 —10 transformants/ug DNA) in Pichia guilliermondii. Based on the PgARS element a series of E. coli—P. guilliermondii shuttle vectors was developed.