["Illegal" transformation of red adenine-dependent mutants of the yeast Pichia pinus by the pYE(ADE2)2 plasmid].
The red adenine-dependent mutants ade1 of the yeast Pichia pinus blocked in the VI step of adenine biosynthesis (lack of AIR-carboxylase) and ade2 mutants blocked in the VII step of adenine biosynthesis (lack of SAIKAR-synthase) were transformed with the plasmid pYE(ADE2)2 containing ADE2 gene of Saccharomyces cerevisiae encoding AIR-carboxylase. The appearance of white Ade+ clones with the frequency 2-7.10(-8) (which is ten-fold higher than reversion frequency) was only observed in the case of ade2 transformation. Genetic analysis points to connection of the "illegitimate" transformants' appearance with the change in the mutant ade2 locus or in a locus closely linked to the former. Ade+ phenotype was stable during 20 generations of mitotic budding. Southern blotting assay of transformant chromosomal DNA indicates that reconstitution of ade2 defective gene is related with its "correction", owing to integration of pYE(ADE2)2 sequence in the vicinity of the mutant locus.
Nestat
M A
MA
Benevolenskiĭ
S V
SV
Dutova
T A
TA
Tolstorukov
I I
II
D000225:Adenine / Q000096:biosynthesis*; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics; D009154:Mutation; D009693:Nucleic Acid Hybridization; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D004718:Saccharomycetales / Q000235:genetics*; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes; D000225:Adenine
"Nezakonnaia" transformatsiia krasnykh adeninzavisimykh mutantov drozhzheĭ Pichia pinus plazmidoĭ pYE(ADE2)2.