Home LiteratureArticle Details
PMID: 16593723 Published · ppublish English Journal Article

Efficient cloning of genes of Neurospora crassa.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 13 ·1986-07-00 ·Pages 4869-73

Vollmer SJ, Yanofsky C

Abstract

We have constructed a genomic library of Neurospora crassa DNA in a cosmid vector that contains the dominant selectable marker for benomyl resistance. The library is arranged to permit the rapid cloning of Neurospora genes by either sib-selection or colony-hybridization protocols. Detailed procedures for the uses of the library are described. By use of these procedures, a modest number of unrelated genes have been isolated. The cloning of trp-3, the structural gene for the multifunctional enzyme tryptophan synthetase (tryptophan synthase, EC 4.2.1.20), is reported in detail; its identity was verified by restriction fragment length polymorphism mapping. The strategies described in this paper should be of use in the cloning of any gene of Neurospora, as well as genes of other lower eukaryotes.

Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Vollmer S J
Department of Biological Sciences, Stanford University, Stanford, CA 94305.
Yanofsky C
References (32)
32 references, click to expand
  1. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I.
    J Mol Biol. 1977 Jun 15;113(1):237-51 PMID: 881736
  2. Studies on transformation of Escherichia coli with plasmids.
    J Mol Biol. 1983 Jun 5;166(4):557-80 PMID: 6345791
  3. In vitro packaging of lambda and cosmid DNA.
    Methods Enzymol. 1979;68:299-309 PMID: 161604
  4. Characterization of the sequence complexity and organization of the Neurospora crassa genome.
    Biochemistry. 1979 Aug 21;18(17):3705-13 PMID: 157775
  5. Expression in Escherichia coli K-12 of the structural gene for catabolic dehydroquinase of Neurospora crassa.
    Proc Natl Acad Sci U S A. 1977 Aug;74(8):3508-12 PMID: 143663
  6. A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome.
    Cell. 1976 Sep;9(1):91-9 PMID: 788919
  7. Regulation of tryptophan biosynthetic enzymes in Neurospora crassa.
    J Bacteriol. 1971 Jul;107(1):193-202 PMID: 5563869
  8. The subunit structure of tryptophan synthase from Neurospora crassa.
    J Biol Chem. 1975 Apr 25;250(8):2941-6 PMID: 123527
  9. FINE STRUCTURE ANALYSIS AT THE TD LOCUS OF NEUROSPORA CRASSA.
    Genetics. 1964 Jan;49:145-58 PMID: 14105106
  10. A GENETIC MAP OF THE TD LOCUS OF NEUROSPORA CRASSA.
    Genetics. 1964 Jan;49:135-44 PMID: 14105105
  11. Studies on the mechanism of the tryptophan synthetase reaction.
    Biochim Biophys Acta. 1962 Aug 13;62:279-93 PMID: 13885361
  12. Transformation of Neurospora crassa with circular and linear DNA and analysis of the fate of the transforming DNA.
    Curr Genet. 1985;10(3):205-12 PMID: 2842071
  13. Cloning and characterization of the multifunctional his-3 gene of Neurospora crassa.
    Gene. 1985;39(2-3):129-40 PMID: 3005109
  14. Plasmid recovery from transformants and the isolation of chromosomal DNA segments improving plasmid replication in Neurospora crassa.
    Curr Genet. 1985;9(5):383-8 PMID: 2967124
  15. Isolation and structural organization of the Neurospora crassa copper metallothionein gene.
    EMBO J. 1985 Oct;4(10):2665-8 PMID: 2932331
  16. General method for cloning Neurospora crassa nuclear genes by complementation of mutants.
    Mol Cell Biol. 1985 Sep;5(9):2272-8 PMID: 2942762
  17. Isolation and characterization of genes differentially expressed during conidiation of Neurospora crassa.
    Mol Cell Biol. 1985 Apr;5(4):849-55 PMID: 3157864
  18. Gene disruption by transformation in Neurospora crassa.
    Mol Cell Biol. 1985 Jul;5(7):1554-9 PMID: 3160929
  19. Identification and chromosomal distribution of 5S rRNA genes in Neurospora crassa.
    Proc Natl Acad Sci U S A. 1985 Apr;82(7):2067-71 PMID: 3157192
  20. Transformation of Neurospora crassa with recombinant plasmids containing the cloned glutamate dehydrogenase (am) gene: evidence for autonomous replication of the transforming plasmid.
    Mol Cell Biol. 1984 Oct;4(10):2041-51 PMID: 6095037
  21. Chromosomal loci of Neurospora crassa.
    Microbiol Rev. 1982 Dec;46(4):426-570 PMID: 6219280
  22. Tryptophan synthase: structure, function, and subunit interaction.
    Adv Enzymol Relat Areas Mol Biol. 1979;49:127-86 PMID: 400853
  23. Organization of the ribosomal ribonucleic acid genes in various wild-type strains and wild-collected strains of Neurospora.
    Mol Gen Genet. 1984;196(2):275-82 PMID: 6092870
  24. Structure of the trifunctional trp-1 gene from Neurospora crassa and its aberrant expression in Escherichia coli.
    J Mol Appl Genet. 1983;2(1):83-99 PMID: 6221060
  25. Cloning of the structural gene for orotidine 5'-phosphate carboxylase of Neurospora crassa by expression in Escherichia coli.
    Mol Gen Genet. 1983;190(3):403-5 PMID: 6308396
  26. Nucleotide sequence of the cloned mRNA and gene of the ADP/ATP carrier from Neurospora crassa.
    EMBO J. 1984 Feb;3(2):377-82 PMID: 6325169
  27. Construction of a shuttle vector for the filamentous fungus Neurospora crassa.
    Proc Natl Acad Sci U S A. 1983 Feb;80(4):1058-62 PMID: 6302667
  28. Rapid and efficient cosmid cloning.
    Nucleic Acids Res. 1981 Jul 10;9(13):2989-98 PMID: 6269067
  29. The construction of cosmid libraries of eukaryotic DNA using the Homer series of vectors.
    Nucleic Acids Res. 1982 Apr 24;10(8):2503-20 PMID: 6281732
  30. Characterization of deletion derivatives of an autonomously replicating Neurospora plasmid.
    Nucleic Acids Res. 1984 Aug 10;12(15):6169-78 PMID: 6236427
  31. Genome organization and characterization of the repetitive and inverted repeat DNA sequences in Neurospora crassa.
    J Biol Chem. 1980 Feb 10;255(3):1138-45 PMID: 6444301
  32. Efficient transformation of Neurospora crassa by utilizing hybrid plasmid DNA.
    Proc Natl Acad Sci U S A. 1979 Oct;76(10):5259-63 PMID: 159454
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.83.13.4869
Published
1986-07-00
Pages
4869-73
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC323844
External Links
PubMed source DOI: 10.1073/pnas.83.13.4869 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2