Current Genetics

, Volume 18, Issue 1, pp 53–58 | Cite as

Distant upstream regulatory sequences control the level of expression of the am (GDH) locus of Neurospora crassa

  • G. D. Frederick
  • J. A. Kinsey
Original Articles
Received:
Accepted:

Summary

We have constructed deletions in the 5′ non-coding sequences of the cloned Neurospora crassa am gene. Vectors with a truncated fragment of the am gene were used in transformation experiments to introduce the deletions into the chromosome by homologous recombination. Analysis of glutamate dehydrogenase (GDH) expression by enzyme assay and immunoblots, as well as Northern and dot blots of poly (A)+ RNA, in the deletion strains indicates that there are two upstream regulatory sequences that control the level of gene expression. The closer of these two elements (URSamα) is at approximately 1.4 kb upstream of the transcriptional start site. The second elements (URSamβ) is located between 2.1 and 3.2 kb upstream of the transcription start site. Deletion of either of these two elements reduces am expression to about 50% of the wild-type level. Deletion of both elements reduce am expression to from 5–16% of the wild-type level. Deletion of 1.1 kb of sequence just downstream of URSamα, which brings this element to within 300 bp of the transcription start site, had no effect on am expression. Likewise, deletion of 3.5 kb of sequence upstream of URSamβ had no effect on expression. None of these deletions had any effect on the expression of usg-1, a gene of unknown function that is transcribed in the same direction as the am gene, and which terminates about 3.5 kb upstream of the URSamβ element.

Key words

Transcription Gene expression In vitro mutagenesis Gene replacement 
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Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • G. D. Frederick
    • 1
  • J. A. Kinsey
    • 1
  1. 1.Department of Microbiology, Molecular Genetics and ImmunologyUniversity of Kansas Medical SchoolKansas CityUSA

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