Screening of growth- or development-related genes by using genomic library with inducible promoter in Aspergillus nidulans

J Microbiol. 2005 Dec;43(6):523-8.

Abstract

Using the genomic library constructed at the downstream of the niiA promoter, which induces the over-expression of an inserted DNA fragment, we have attempted to screen the genes affecting growth or development by over-expression. The wild-type strain was transformed using the AMA-niiA(p) library and cultured on 1.2 M sorbitol media, in which asexual sporulation is induced, but sexual development is repressed. Over 100,000 strains transformed to pyrG(+) were analyzed with regard to any changes in phenotype. Consequently, seven strains were isolated for further analyses. These strains were designated NOT [niiA(p) over-expression transformants] stains. Four of the strains were of the inducible type, and the remaining strains were of the multi-copy suppression type. Two of the inducible-type strains, NOT1 and NOT40, harbored genes which had been inserted in reverse direction, suggesting that the mutant phenotypes had been derived from an excess amount of anti-sense mRNA. Domain analyses of the deduced polypeptides from the DNA fragments rescued from the transformants revealed that NOT1, NOT40 and NOT6 harbored a LisH motif, a forkhead domain, and a Zn(II)(2)Cys(6) binuclear zinc cluster, respectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aspergillus nidulans / cytology
  • Aspergillus nidulans / genetics*
  • Aspergillus nidulans / growth & development
  • Conserved Sequence
  • Gene Dosage
  • Gene Expression
  • Gene Library
  • Genes, Fungal*
  • Genetic Vectors
  • Molecular Biology / methods*
  • Molecular Sequence Data
  • Morphogenesis / genetics
  • Promoter Regions, Genetic
  • Protein Structure, Tertiary / genetics
  • RNA, Antisense
  • Sequence Alignment

Substances

  • RNA, Antisense