Cloning and physical characterization of linked lysine genes (lys4, lys15) of Saccharomyces cerevisiae.
The plasmid pSC4 which carries a 7.8 kb yeast DNA insert at the BamHI site of the Vector YEp13, complemented simultaneously MO-59-13c lys4, LU75 lys15 and LU32 lys4lys15 (double) mutations of Saccharomyces cerevisiae. The 1.9 kb BamHI-XbaI DNA insert of the subclone pSO51 complemented the LU75 lys15 mutation. The 2.8 kb Xhol-XhoI DNA insert of the pSO52 subclone, like pSC4, complemented all three mutations. The 1.9 kb BamHI-XbaI DNA and the 2.8 kb Xhol-XhoI DNA were 100 bp apart in the pSC4 DNA insert and exhibited no homology with each other upon Southern hybridization. The 1.9 kb BamHI-XbaI DNA insert exhibited homology with the pSC4 and pSO51 DNA as well as the genomic DNA of MO-59-13c lys4, LU75 lys15, LU32 lys4lys15, and RC1 (LYS) when digested with appropriate restriction enzymes. The 2.8 kb XhoI-XhoI DNA insert exhibited homology with the pSC4 and pSO52 DNA as well as MO-59-13c lys4, LU75 lys15, LU32 lys4lys15, and RC1 (LYS) genomic DNA, when digested with XhoI enzyme. The 2.8 kb DNA probe also hybridized with ply(A)+ RNA from RC1 and lys4+ transformant but not that from MO-59-13c lys4 mutant.
Wang
L
L
Department of Microbiology, Miami University, Oxford, OH 45056.
Okamoto
S
S
Bhattacharjee
J K
JK
D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D008040:Genetic Linkage; D008239:Lysine / Q000096:biosynthesis / Q000235:genetics*; D009154:Mutation; D010957:Plasmids; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014158:Transcription, Genetic; D014170:Transformation, Genetic
D004271:DNA, Fungal; D008239:Lysine