[A novel approach to isolation and functional characterization of genomic DNA from the methylotrophic yeast Hansenula polymorpha].
A novel approach to isolation and functional characterization of the Hansenula polymorpha genes basing on the use of two strains of different origin is described. One of these strains is better suited for the isolation of genomic DNA fragments, while the other is preferable for their functional analysis. Thirty three genomic sequences governing expression of a reporter protein have been isolated. Analysis of the sequence encoding a homolog of the Saccharomyces cerevisiae cofilin revealed two introns. Another isolated DNA fragment encoded a homolog of the S. cerevisiae V ps10p. Disruption of the corresponding gene resulted in secretion of a vacuolar protein, carboxypeptidase Y, into the culture medium.
Agafonov
M O
MO
Institute of Experimental Cardiology, Cardiology Research Center, Moscow, 121552 Russia. aga@cardio.ru
Deev
A V
AV
Kim
S Y
SY
Sohn
J H
JH
Choi
E S
ES
Ter-Avanesian
M D
MD
D051339:Actin Depolymerizing Factors; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004271:DNA, Fungal / Q000302:isolation & purification*; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D016678:Genome; D007438:Introns; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D021381:Protein Transport / Q000235:genetics; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014617:Vacuoles / Q000378:metabolism; D033921:Vesicular Transport Proteins
D051339:Actin Depolymerizing Factors; D004271:DNA, Fungal; D005656:Fungal Proteins; D008840:Microfilament Proteins; C087335:PEP1 protein, S cerevisiae; D011956:Receptors, Cell Surface; D029701:Saccharomyces cerevisiae Proteins; D033921:Vesicular Transport Proteins