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Mol Cell Biol. Jul 1987; 7(7): 2506–2511.
PMCID: PMC365384

Molecular cloning and characterization of the cys-3 regulatory gene of Neurospora crassa.

Abstract

The regulatory gene cys-3+ controls the synthesis of a number of enzymes involved in sulfur metabolism. cys-3 mutants show a multiple loss of enzymes in different pathways of sulfur metabolism. The cys-3+ gene was isolated by transformation of an aro-9 qa-2 cys-3 inl strain with a clone bank followed by screening with the "sib selection" method. The library used (pRAL1) contained inserts of Sau3a partial digest fragments of about 9 kilobases as well as the Neurospora qa-2+ gene. Double selection for qa-2+ and cys-3+ function was carried out. The transformants obtained with the isolated cys-3+ clone show recovery of the enzyme activities associated with the cys-3 mutation (e.g., arylsulfatase and sulfate permease). Restriction fragment length polymorphism experiments confirmed the identity of the clone, mRNA studies with Northern blots show that the expression of the cys-3+ gene is inducible. In contrast to cys-3+, the cys-3 (P22) mutant gene was not expressed at a higher level under sulfur-derepressed conditions.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Akins RA, Lambowitz AM. General method for cloning Neurospora crassa nuclear genes by complementation of mutants. Mol Cell Biol. 1985 Sep;5(9):2272–2278. [PMC free article] [PubMed]
  • Aviv H, Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. [PMC free article] [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Dagert M, Ehrlich SD. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. [PubMed]
  • Dietrich PS, Metzenberg RL. Metabolic suppressors of a regulatory mutant in Neurospora. Biochem Genet. 1973 Jan;8(1):73–84. [PubMed]
  • Marzluf GA. Genetic and metabolic controls for sulfate metabolism in Neurospora crassa: isolation and study of chromate-resistant and sulfate transport-negative mutants. J Bacteriol. 1970 Jun;102(3):716–721. [PMC free article] [PubMed]
  • Marzluf GA. Genetic and metabolic control of sulfate metabolism in Neurospora crassa: a specific permease for choline-O-sulfate. Biochem Genet. 1972 Dec;7(3):219–233. [PubMed]
  • Marzluf GA, Metzenberg RL. Positive control by the cys-3 locus in regulation of sulfur metabolism in Neurospora. J Mol Biol. 1968 Apr 28;33(2):423–437. [PubMed]
  • Metzenberg RL, Ahlgren SK. Mutants of Neurospora deficient in aryl sulfatase. Genetics. 1970 Mar-Apr;64(3):409–422. [PMC free article] [PubMed]
  • Paietta J, Marzluf GA. Plasmid recovery from transformants and the isolation of chromosomal DNA segments improving plasmid replication in Neurospora crassa. Curr Genet. 1985;9(5):383–388. [PubMed]
  • Paietta JV, Marzluf GA. Gene disruption by transformation in Neurospora crassa. Mol Cell Biol. 1985 Jul;5(7):1554–1559. [PMC free article] [PubMed]
  • Pall ML. Amino acid transport in Neurospora crassa. IV. Properties and regulation of a methionine transport system. Biochim Biophys Acta. 1971 Mar 9;233(1):201–214. [PubMed]
  • Perkins DD, Radford A, Newmeyer D, Björkman M. Chromosomal loci of Neurospora crassa. Microbiol Rev. 1982 Dec;46(4):426–570. [PMC free article] [PubMed]
  • Reinert WR, Patel VB, Giles NH. Genetic regulation of the qa gene cluster of Neurospora crassa: induction of qa messenger ribonucleic acid and dependency on qa-1 function. Mol Cell Biol. 1981 Sep;1(9):829–835. [PMC free article] [PubMed]
  • Rigby PW, Dieckmann M, Rhodes C, Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Stohl LL, Lambowitz AM. Construction of a shuttle vector for the filamentous fungus Neurospora crassa. Proc Natl Acad Sci U S A. 1983 Feb;80(4):1058–1062. [PMC free article] [PubMed]

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