The orotidine-5′-phosphate decarboxylase gene (URA3) of a methylotrophic yeast, Candida boidinii: Nucleotide sequence and its expression in Escherichia coli

High-frequency transformation of a methylotrophic yeast, Candida boidinii, with autonomously replicating plasmids which are also functional in Saccharomyces cerevisiae

“…Evidently, C. boidindi URA3 DNA itself contains an ARS activity for S. cerevisiae. We analyzed the C. boidindi URA3 sequence (34) and found that a single region (-948 to -987 on the complementary strand [numbering from the initiator ATG]) was homologous to both the ARS core sequence (ATATATAYIT, 10 of 11 match) and the ARS box sequence ('1TIlGAA, 5 of 6 match), both of which were reported to be essential for S. cerevisiae ARS activity (6,29).…”

“…S. cerevisiae YPH500 was transformed with plasmids pBARCU1, pBARCU2, and pBCU351 (34). All of these plasmids could transform S. cerevisiae YPH500 to URA+ phenotype at high frequencies of 7.5 x 104, 6.1 x 104, and 4.5 x 104 CFU/,ug of DNA, respectively (Table 1).…”

“…At first, C. boidindi genomic DNA was completely digested with EcoRI or HindIII, ligated into the corresponding unique site of plasmid pBCU351 (34), and introduced into E. coli JM109 cells. A total of 1.8 x 104 independent colonies were obtained for the EcoRI Transformation of C. boidinii TK62 with pBARCU1 and pBARCU2 by a modified lithium acetate method.…”

Isolation and sequence analysis of the orotidine-5'-phosphate decarboxylase gene (URA3) ofCandida utilis. Comparison with the OMP decarboxylase gene family

“…Absolutely conserved residues are marked with (*) and high conserved residues with (.). The sequences are: DCOP_CANUT from C. utilis (this study), DCOP_CANPA from C. parapsilosis (Nosek, 1996b), DCOP_CANMA from Candida maltosa (Ohkuma et al, 1993), DCOP_CANAL from Candida albicans (Ernst and Losberger, 1989), DCOP_CANBO from Candida boidinii (Sakai et al, 1992), DCOP_CANGA from Candida glabratra (Zhou et al, 1994), DCOP_CANTR from Candida tropicalis (Cregg et al, 1990), DCOP_KLUMA from K. marxianus (Bergkamp et al, 1993), DCOP_KLULA from Kluyveromyces lactis (Shuster et al, 1987), DCOP_SCERV from S. cerevisiae (Rose et al, 1984), DCOP_HANPO from Hansenula polymorpha (Merckelbach et al, 1993), DCOP_HANAN from Hansenula anomala (Ogata et al, 1992), DCOP_HANFA from H. fabianii (M. Kato et al, unpublished results), DCOP_PICST from P. stipitis (Yang et al, 1994), DCOP_PICOH from P. ohmeri (Piredda and Gaillardin, 1994), DCOP_PICAN from Pichia anomala (Pérez et al, 1996), DCOP_YARLY from Yarrowia lipolytica (Strick et al, 1995), DCOP_PHYBL from Phycomyces blakesleeanus (Díaz-Minguez et al, 1990), DCOP_RHYCI from Rhyzomucor circinelloides (Benito et al, 1992), DCOP_SCHPO from S. pombe (Grimm et al, 1988), DCOP_SCHCO from Schizophyllum commune (Froeliger et al, 1989), DCOP_PENCH from Penicillium chrysogenum (Cantoral et al, 1988), DCOP_MOUSE from Mus musculus (mouse) (Ohmstede et al, 1986), PYR5_HUMAN from Homo sapiens (human) (Suttle et al, 1988), PYR5_BOVIN from Bos taurus (bovine) (Shoeberg et al, 1993), DCOP_ASPNG from Aspergillus niger (Wilson et al, 1988), DCOP_EMENI from Aspergillus nidulans (Oakley et al, 1987), DCOP_SORMA from Sordaria macrospora (Nowrousian, 1996), DCOP_USTMA from Ustilago maydis (Kronstad et al, 1989), DCOP_NICTA from Nicotiana tabacum (Millar and Kunst, 1995), DCOP_ARATH from Arabidopsis thaliana …”

Directed mutagenesis in an asporogenous methylotrophic yeast: cloning, sequencing, and one-step gene disruption of the 3-isopropylmalate dehydrogenase gene (LEU2) of Candida boidinii to derive doubly auxotrophic marker strains

“…The C. boidinii LEU2 disruption vectors pLS, pLSB, and pLSP were constructed as follows. The 654-bp SalI-BamHI fragment (the SB fragment) and the 1,049-bp PstI-SalI fragment (the SP fragment) were parts of the 3,612-bp Sall fragment harboring the C. boidinii URA3 gene (22,23). The SB and SP fragments were each ligated with the 3.6-kb Sall C. boidinii URA3 fragment, and the fragments obtained (4.3 and 4.7 kb, respectively) were cloned into the multiple cloning site of pBluescriptll SK+ (pSBR for the SB fragment, pSPR for the SP fragment).…”