Isolation by genetic complementation of two differentially expressed genes for β-isopropylmalate dehydrogenase from Aspergillus niger

Williams, B. A. ; Sillaots, S. ; Tsang, A. ; Storms, R.
Springer
Published 1996
ISSN:
1432-0983
Keywords:
Key words Aspergillus niger ; Leucine biosynthesis ; Genetic complementation ; Differential expression
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract  We have constructed an Aspergillus niger cDNA library with a yeast expression vector. The library DNA complemented a leucine auxotroph of Saccharomyces cerevisiae (strain BWG1-7a) at a frequency of 4×10-4. Plasmids rescued from the yeast prototrophs also complemented Escherichia coli (strain MC1066) deficient in leucine biosynthesis. Sequence determination of the rescued plasmids revealed two genes for β-isopropylmalate dehydrogenase, which we called leu2A and leu2B. Genomic-blot analysis suggested that both leu2A and leu2B were derived from single-copy genes. Northern-blot hybridization showed that in nutrient-rich medium a leu2A transcript accumulated during germination and log-phase growth while the leu2B transcript appeared late in the growth phase. In minimal medium, only leu2A expression was greatly stimulated. We examined the codon preference of these two genes. Whereas leu2A shows a bias in codon usage typical of A. niger genes, leu2B does not. These results indicate the presence in A. niger of two highly divergent, differentially regulated, isozymes for β-isopropylmalate dehydrogenase.
Type of Medium:
Electronic Resource
URL:
_version_ 1798295559401897984
autor Williams, B. A.
Sillaots, S.
Tsang, A.
Storms, R.
autorsonst Williams, B. A.
Sillaots, S.
Tsang, A.
Storms, R.
book_url http://dx.doi.org/10.1007/s002940050137
datenlieferant nat_lic_papers
hauptsatz hsatz_simple
identnr NLM203129326
issn 1432-0983
journal_name Current genetics
materialart 1
notes Abstract  We have constructed an Aspergillus niger cDNA library with a yeast expression vector. The library DNA complemented a leucine auxotroph of Saccharomyces cerevisiae (strain BWG1-7a) at a frequency of 4×10-4. Plasmids rescued from the yeast prototrophs also complemented Escherichia coli (strain MC1066) deficient in leucine biosynthesis. Sequence determination of the rescued plasmids revealed two genes for β-isopropylmalate dehydrogenase, which we called leu2A and leu2B. Genomic-blot analysis suggested that both leu2A and leu2B were derived from single-copy genes. Northern-blot hybridization showed that in nutrient-rich medium a leu2A transcript accumulated during germination and log-phase growth while the leu2B transcript appeared late in the growth phase. In minimal medium, only leu2A expression was greatly stimulated. We examined the codon preference of these two genes. Whereas leu2A shows a bias in codon usage typical of A. niger genes, leu2B does not. These results indicate the presence in A. niger of two highly divergent, differentially regulated, isozymes for β-isopropylmalate dehydrogenase.
package_name Springer
publikationsjahr_anzeige 1996
publikationsjahr_facette 1996
publikationsjahr_intervall 8004:1995-1999
publikationsjahr_sort 1996
publisher Springer
reference 30 (1996), S. 305-311
schlagwort Key words Aspergillus niger
Leucine biosynthesis
Genetic complementation
Differential expression
search_space articles
shingle_author_1 Williams, B. A.
Sillaots, S.
Tsang, A.
Storms, R.
shingle_author_2 Williams, B. A.
Sillaots, S.
Tsang, A.
Storms, R.
shingle_author_3 Williams, B. A.
Sillaots, S.
Tsang, A.
Storms, R.
shingle_author_4 Williams, B. A.
Sillaots, S.
Tsang, A.
Storms, R.
shingle_catch_all_1 Williams, B. A.
Sillaots, S.
Tsang, A.
Storms, R.
Isolation by genetic complementation of two differentially expressed genes for β-isopropylmalate dehydrogenase from Aspergillus niger
Key words Aspergillus niger
Leucine biosynthesis
Genetic complementation
Differential expression
Key words Aspergillus niger
Leucine biosynthesis
Genetic complementation
Differential expression
Abstract  We have constructed an Aspergillus niger cDNA library with a yeast expression vector. The library DNA complemented a leucine auxotroph of Saccharomyces cerevisiae (strain BWG1-7a) at a frequency of 4×10-4. Plasmids rescued from the yeast prototrophs also complemented Escherichia coli (strain MC1066) deficient in leucine biosynthesis. Sequence determination of the rescued plasmids revealed two genes for β-isopropylmalate dehydrogenase, which we called leu2A and leu2B. Genomic-blot analysis suggested that both leu2A and leu2B were derived from single-copy genes. Northern-blot hybridization showed that in nutrient-rich medium a leu2A transcript accumulated during germination and log-phase growth while the leu2B transcript appeared late in the growth phase. In minimal medium, only leu2A expression was greatly stimulated. We examined the codon preference of these two genes. Whereas leu2A shows a bias in codon usage typical of A. niger genes, leu2B does not. These results indicate the presence in A. niger of two highly divergent, differentially regulated, isozymes for β-isopropylmalate dehydrogenase.
1432-0983
14320983
Springer
shingle_catch_all_2 Williams, B. A.
Sillaots, S.
Tsang, A.
Storms, R.
Isolation by genetic complementation of two differentially expressed genes for β-isopropylmalate dehydrogenase from Aspergillus niger
Key words Aspergillus niger
Leucine biosynthesis
Genetic complementation
Differential expression
Key words Aspergillus niger
Leucine biosynthesis
Genetic complementation
Differential expression
Abstract  We have constructed an Aspergillus niger cDNA library with a yeast expression vector. The library DNA complemented a leucine auxotroph of Saccharomyces cerevisiae (strain BWG1-7a) at a frequency of 4×10-4. Plasmids rescued from the yeast prototrophs also complemented Escherichia coli (strain MC1066) deficient in leucine biosynthesis. Sequence determination of the rescued plasmids revealed two genes for β-isopropylmalate dehydrogenase, which we called leu2A and leu2B. Genomic-blot analysis suggested that both leu2A and leu2B were derived from single-copy genes. Northern-blot hybridization showed that in nutrient-rich medium a leu2A transcript accumulated during germination and log-phase growth while the leu2B transcript appeared late in the growth phase. In minimal medium, only leu2A expression was greatly stimulated. We examined the codon preference of these two genes. Whereas leu2A shows a bias in codon usage typical of A. niger genes, leu2B does not. These results indicate the presence in A. niger of two highly divergent, differentially regulated, isozymes for β-isopropylmalate dehydrogenase.
1432-0983
14320983
Springer
shingle_catch_all_3 Williams, B. A.
Sillaots, S.
Tsang, A.
Storms, R.
Isolation by genetic complementation of two differentially expressed genes for β-isopropylmalate dehydrogenase from Aspergillus niger
Key words Aspergillus niger
Leucine biosynthesis
Genetic complementation
Differential expression
Key words Aspergillus niger
Leucine biosynthesis
Genetic complementation
Differential expression
Abstract  We have constructed an Aspergillus niger cDNA library with a yeast expression vector. The library DNA complemented a leucine auxotroph of Saccharomyces cerevisiae (strain BWG1-7a) at a frequency of 4×10-4. Plasmids rescued from the yeast prototrophs also complemented Escherichia coli (strain MC1066) deficient in leucine biosynthesis. Sequence determination of the rescued plasmids revealed two genes for β-isopropylmalate dehydrogenase, which we called leu2A and leu2B. Genomic-blot analysis suggested that both leu2A and leu2B were derived from single-copy genes. Northern-blot hybridization showed that in nutrient-rich medium a leu2A transcript accumulated during germination and log-phase growth while the leu2B transcript appeared late in the growth phase. In minimal medium, only leu2A expression was greatly stimulated. We examined the codon preference of these two genes. Whereas leu2A shows a bias in codon usage typical of A. niger genes, leu2B does not. These results indicate the presence in A. niger of two highly divergent, differentially regulated, isozymes for β-isopropylmalate dehydrogenase.
1432-0983
14320983
Springer
shingle_catch_all_4 Williams, B. A.
Sillaots, S.
Tsang, A.
Storms, R.
Isolation by genetic complementation of two differentially expressed genes for β-isopropylmalate dehydrogenase from Aspergillus niger
Key words Aspergillus niger
Leucine biosynthesis
Genetic complementation
Differential expression
Key words Aspergillus niger
Leucine biosynthesis
Genetic complementation
Differential expression
Abstract  We have constructed an Aspergillus niger cDNA library with a yeast expression vector. The library DNA complemented a leucine auxotroph of Saccharomyces cerevisiae (strain BWG1-7a) at a frequency of 4×10-4. Plasmids rescued from the yeast prototrophs also complemented Escherichia coli (strain MC1066) deficient in leucine biosynthesis. Sequence determination of the rescued plasmids revealed two genes for β-isopropylmalate dehydrogenase, which we called leu2A and leu2B. Genomic-blot analysis suggested that both leu2A and leu2B were derived from single-copy genes. Northern-blot hybridization showed that in nutrient-rich medium a leu2A transcript accumulated during germination and log-phase growth while the leu2B transcript appeared late in the growth phase. In minimal medium, only leu2A expression was greatly stimulated. We examined the codon preference of these two genes. Whereas leu2A shows a bias in codon usage typical of A. niger genes, leu2B does not. These results indicate the presence in A. niger of two highly divergent, differentially regulated, isozymes for β-isopropylmalate dehydrogenase.
1432-0983
14320983
Springer
shingle_title_1 Isolation by genetic complementation of two differentially expressed genes for β-isopropylmalate dehydrogenase from Aspergillus niger
shingle_title_2 Isolation by genetic complementation of two differentially expressed genes for β-isopropylmalate dehydrogenase from Aspergillus niger
shingle_title_3 Isolation by genetic complementation of two differentially expressed genes for β-isopropylmalate dehydrogenase from Aspergillus niger
shingle_title_4 Isolation by genetic complementation of two differentially expressed genes for β-isopropylmalate dehydrogenase from Aspergillus niger
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wilbert
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source_archive Springer Online Journal Archives 1860-2000
timestamp 2024-05-06T09:38:08.250Z
titel Isolation by genetic complementation of two differentially expressed genes for β-isopropylmalate dehydrogenase from Aspergillus niger
titel_suche Isolation by genetic complementation of two differentially expressed genes for β-isopropylmalate dehydrogenase from Aspergillus niger
topic W
uid nat_lic_papers_NLM203129326