Use of Gene Replacement Transformation to Elucidate Gene Function in the Qa Gene Cluster of Neurospora Crassa

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RESUMO

Gene replacement by transformation, employing selective genetic recombination techniques, has been used to delete or disrupt the qa-x, qa-y and qa-1S genes of the qa gene cluster of Neurospora crassa. The growth characteristics of the strain carrying the deletion of the qa-y gene support earlier evidence that this gene encodes a quinic acid permease. The strain containing the deletion of the qa-1S gene (Δqa-1S) was examined with respect to quinic acid induction and carbon catabolite repression. The Δqa-1S strain exhibits constitutive expression of the qa genes supporting earlier evidence that the qa-1S gene codes for a repressor. Several of the qa genes continued to be expressed at high levels even in the presence of glucose in the Δqa-1S strain, which indicates that transcription of these genes is not being affected directly by a repressor molecule in the presence of glucose.

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