Tagging Hansenula polymorpha genes by random integration of linear DNA fragments (RALF)
نویسندگان
چکیده
منابع مشابه
Pexophagy in Hansenula polymorpha.
In the yeast Hansenula polymorpha the development and turnover of peroxisomes is readily achieved by manipulation of the cultivation conditions. The organelles massively develop when the cells are incubated in the presence of methanol as the sole source of carbon and energy. However, they are rapidly and selectively degraded when methanol-grown cells are placed at conditions of repression of me...
متن کاملMethanol oxidation and assimilation in Hansenula polymorpha
The metabolism of methanol was monitored in whole cells of the methylotrophic yeast Hansenula polymorpha by using [13C]methanol and n.m.r. in vivo. The main products observed under normal conditions were trehalose and glycerol, whereas cells that were starved before exposure to [13C]methanol also accumulated glutamate, glutamine and alanine; formate was also more prominent in spectra from starv...
متن کاملHansenula polymorpha Vam7p is required for macropexophagy.
We have analyzed the functions of two vacuolar t-SNAREs, Vam3p and Vam7p, in peroxisome degradation in the methylotrophic yeast Hansenula polymorpha. A Hp-vam7 mutant was strongly affected in peroxisome degradation by selective macropexophagy as well as non-selective microautophagy. Deletion of Hp-Vam3p function had only a minor effect on peroxisome degradation processes. Both proteins were loc...
متن کاملTagging developmental genes in Dictyostelium by restriction enzyme-mediated integration of plasmid DNA.
Introduction of restriction enzyme along with linearized plasmid results in integration of plasmid DNA at genomic restriction sites in a high proportion of the resulting transformants. We have found that electroporating BamHI or EcoRI together with pyr5-6 plasmids cut with the same enzyme stimulates the efficiency of transformation in Dictyostelium discoideum more than 20-fold over the rate see...
متن کاملChromosomal targeting of replicating plasmids in the yeast Hansenula polymorpha.
Using an optimized transformation protocol we have studied the possible interactions between transforming plasmid DNA and the Hansenula polymorpha genome. Plasmids consisting only of a pBR322 replicon, an antibiotic resistance marker for Escherichia coli and the Saccharomyces cerevisiae LEU2 gene were shown to replicate autonomously in the yeast at an approximate copy number of 6 (copies per ge...
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ژورنال
عنوان ژورنال: Molecular Genetics and Genomics
سال: 2001
ISSN: 1617-4615,1617-4623
DOI: 10.1007/s004380100584