1887

Abstract

pyruvate kinase gene () was transformed into yeast using the multicopy vector pJDB207. Growth rates and gene expression levels varied considerably amongst the transformants. Yeast transformants expressing the gene at high levels formed small colonies compared with those expressing the gene at relatively low levels. Slow-growing transformants reverted at high frequency to more rapid growth, and this correlated with decreases in gene copy number and mRNA abundance. This apparent selection against over-expression was disrupted by the introduction of a stop codon at the 5′-end of the coding region, thus confirming that the growth effects were mediated by the gene. However, massive overproduction of pyruvate kinase in yeast, using multiple copies of a : gene fusion, had no significant effect upon cell growth. This suggests that the deleterious effect upon the host yeast cell is mediated by abnormally high levels of the wild-type gene or mRNA, rather than by increased pyruvate kinase levels.

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1990-12-01
2021-12-01
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