A Neurospora crassa library, constructed in a derivative of the plasmid pBR322 (pRK9), was used to transform two E. coli chlD molybdenum cofactor mutants (chlD, chlD: :Mu). Subsequently, one transformant from each of three independent transformation experiments was restriction mapped. All three tran
Cloning and sequence analysis of the glucoamylase gene ofNeurospora crassa
β Scribed by P. J. Stone; A. J. Makoff; J. H. Parish; A. Radford
- Publisher
- Springer-Verlag
- Year
- 1993
- Tongue
- English
- Weight
- 677 KB
- Volume
- 24
- Category
- Article
- ISSN
- 0172-8083
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β¦ Synopsis
A 1.0-kb DNA fragment, corresponding to an internal region of the Neurospora crassa glucoamylase gene, gla-1, was generated from genomic DNA by the polymerase chain reaction, using oligonucleotide primers which had been deduced from the known N-terminal amino-acid sequence or from consensus regions within the aligned amino,acid sequences of other fungal glucoamylases. The fragment was used to screen an N. crassa genomic DNA library. One clone contained the gene together with flanking regions and its sequence was determined. The gene was found to code for a preproprotein of 626 amino acids, 35 of which constitute a signal and propeptide region. The protein and the gene are compared with corresponding sequences in other fungi.
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