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Wild-type and mutant alleles of the Aspergillus nidulans developmental regulator gene brlA: correlation of variant sites with protein function
Authors:Griffith G W  Stark M S  Clutterbuck A J
Affiliation:(1) Division of Molecular Genetics, Institute of Biomedical and Life Sciences, Pontecorvo Building, Glasgow University, Glasgow G11 6NU, Scotland, UK E-mail: j.clutterbuck@bio.gla.ac.uk Tel.: +44-141-3305105; Fax: +44-141-3304878, GB
Abstract:
The DNA sequences of two wild-type and eleven mutant alleles of the developmental regulator gene brlA from Aspergillus nidulans, which encodes a zinc-finger protein, were characterized. Variant sites were located on rescued plasmids or PCR products based either on their meiotic map position or the use of denaturing gradient gel electrophoresis. Mutations in three null mutants, one of which is partially suppressible, encode premature stop codons. Two environmentally sensitive mutants were characterised by substitution of leucines required for stabilisation of α-helices in each of the two putative zinc-finger domains. A third zinc-finger substitution is predicted to disrupt recognition of a guanine residue in the DNA target. The mutations in four other leaky mutants map C-terminal to the zinc fingers; one minimally leaky mutant has a premature stop codon, which results in the removal of the last 38 residues of the protein product. Received: 16 February 1999 / Accepted: 22 July 1999
Keywords:brlA mutants  Denaturing gradient gel electrophoresis (DGGE)  Suppressible mutant  Mutant site detection  Zinc finger protein
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