There are about 17 chromosomes in yeast Saccharomycescerevisiae.A middle sized chromosome,chromosome V,waschosen in this work for studying and constructing the physi-cal maps.Chromosome V from strain A364a was isolate...There are about 17 chromosomes in yeast Saccharomycescerevisiae.A middle sized chromosome,chromosome V,waschosen in this work for studying and constructing the physi-cal maps.Chromosome V from strain A364a was isolatedby pulsed-field gradient gel electrophoresis(PFGE).Gelslices containing chromosome V DNA were digestedwith two rare cutting enzymes,NotⅠand SfiⅠ,and three6-Nt recognizing enzymes,SmaⅠ,SstⅡ and ApaⅠ.Several strategies-partial or complete digestions,digestion with different sets of two enzymes,and hybrid-ization with cloned genetically mapped probes(CAN1,URA3,CEN5,PRO3,CHO1,SUP19,RAD51,RAD3)——were used to align the restriction fragments.There are 9,9,15,17,and 20 sites for NotⅠ,SfiⅠ,SmaⅠ,SstⅡ and ApaⅠrespectively in the map of the A364a chromosome V.Itstotal length was calculated to be 620 Kb(Kilo-bases).Thedistributions of the cutting sites for these five enzymesthrough the whole chromosome are not uniform.A comp-arison between the physical map and the genetic map wasalso made.展开更多
文摘There are about 17 chromosomes in yeast Saccharomycescerevisiae.A middle sized chromosome,chromosome V,waschosen in this work for studying and constructing the physi-cal maps.Chromosome V from strain A364a was isolatedby pulsed-field gradient gel electrophoresis(PFGE).Gelslices containing chromosome V DNA were digestedwith two rare cutting enzymes,NotⅠand SfiⅠ,and three6-Nt recognizing enzymes,SmaⅠ,SstⅡ and ApaⅠ.Several strategies-partial or complete digestions,digestion with different sets of two enzymes,and hybrid-ization with cloned genetically mapped probes(CAN1,URA3,CEN5,PRO3,CHO1,SUP19,RAD51,RAD3)——were used to align the restriction fragments.There are 9,9,15,17,and 20 sites for NotⅠ,SfiⅠ,SmaⅠ,SstⅡ and ApaⅠrespectively in the map of the A364a chromosome V.Itstotal length was calculated to be 620 Kb(Kilo-bases).Thedistributions of the cutting sites for these five enzymesthrough the whole chromosome are not uniform.A comp-arison between the physical map and the genetic map wasalso made.
