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
Molecular cloning and characterization ofalcthe gene encoding allantoicase ofNeurospora crassa
β Scribed by Hakjoo Lee; Ying-Hui Fu; George A. Marzluf
- Publisher
- Springer
- Year
- 1990
- Tongue
- English
- Weight
- 819 KB
- Volume
- 222
- Category
- Article
- ISSN
- 1617-4615
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β¦ Synopsis
Purines can be utilized as a secondary nitrogen source by Neurospora crassa during conditions of nitrogen limitation. The expression of purine catabolic enzymes is governed by the nitrogen regulatory circuit and requires induction by uric acid. The major positive-acting nitrogen regulatory gene, nit-2, turns on the expression of the purine catabolic enzymes, which may also be subject to negative regulation by a second control gene, nmr. We have cloned alc, the structural gene which encodes allantoicase, an inducible enzyme of the purine degradative pathway. The identity of the alc clone was confirmed by restriction fragment length polymorphism analysis and by repeat-induced mutation. The alc gene is transcribed to give a single messenger RNA, approximately 1.2 kb in length. The negative-acting nmr gene affects the expression of alc in the expected manner. Both the nit-2 and the nmr control genes affect alc mRNA levels and allantoicase enzyme activity in both the induced and nitrogen-repressed conditions.
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Expression of the structural genes of the nitrogen control circuit of Neurospora crassa is regulated by the positive-acting nit-2 control gene and by the negative-acting nmr control gene. Nitrate reductase is expressed in a constitutive fashion in nmr mutant strains, which appear to be largely insen
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