Isolasi dan karakterisasi mutan sal4 di ragi (Saccharomyces cereviceae)
1995
Puspaningsih, Ni Nyoman Tri
Recently, genetics manipulation in yeast Saccharomyces cereviceae have much been done. It because yeast can be used as a host cell alternative in the forign protein expression, therefore information about fidelity from yeast should be studied. Preliminary study showed that sal4 gene has assumed to has a role in translation fidelity control and/or termination factor. To study the gene function, mutation in yeast BSC483/1a has been done by Ethylmethane sulphonate. Mutants wished are mutated at sal4 locus and have characteristic of both allosuppressor and omnipotent suppressor. Phenotype of alosuppresor mutants were indicated by white colour consistency in YPD and Y8 medium, temperature sensitivity, paremomcyn sensitivity and growth rate. Quantitatively, effectiveness as omnipotent suppressor has been done by using gene fuion between PGK and β-galaktosidase. The result showed that BSC483/1a strain could be mutated by Ethylmethane sulphonate 1% and produced eight allosuppressor mutants. Two of them (Number 8 and 10) have characteristic of temperature sensitivity, and the two others (Number 1 and 13) were mutated at sal4 gene locus. Characterize of sal4 mutants (1 and 13) didn't show temperature sensitive and have growth rate re;atively more slowly than the wild type. Mutant (number 13) could suppress nonsense mutation (realthrough) at termination codon UAG with β-galaktosidase activity as amount 2.70 unit/ml.
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