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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
|
| pubmed:dateCreated |
1991-4-22
|
| pubmed:abstractText |
A gene, amy, encoding an alpha-amylase, was cloned on a 4.8 kb Sau3A fragment from the DNA of Streptomyces griseus IMRU3570. The gene was localized to a 2.27 kb fragment by subcloning and deletion mapping experiments. The gene contained an open reading frame (ORF) of 1698 nucleotides that encoded a protein of 566 amino acids with a deduced Mr of 59713 Da. Dot-blot analysis revealed that the copy number of the transcript in S. lividans transformed with the amy gene was 2.8-fold higher than in the donor S. griseus strain in good agreement with the proportionally higher secretion of amylase in S. lividans. A transcription initiation site was found approximately 64 bp upstream from the ATG translation start codon. The promoter of the amy gene was subcloned on a 290 bp HindIII--EcoRI fragment. Expression of a neomycin resistance gene from the amy promoter was negatively regulated by glucose. A 219 nucleotide fragment extending from the single BstEII site to the end of the amy gene was dispensable since active alpha-amylase was secreted after deletion of this region and coupling of a TGA translation stop codon.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Feb
|
| pubmed:issn |
0026-8925
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
225
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
278-88
|
| pubmed:dateRevised |
2008-11-21
|
| pubmed:meshHeading |
pubmed-meshheading:1900915-Amino Acid Sequence,
pubmed-meshheading:1900915-Base Sequence,
pubmed-meshheading:1900915-Blotting, Northern,
pubmed-meshheading:1900915-Cloning, Molecular,
pubmed-meshheading:1900915-Drug Resistance,
pubmed-meshheading:1900915-Gene Expression,
pubmed-meshheading:1900915-Genes, Bacterial,
pubmed-meshheading:1900915-Molecular Sequence Data,
pubmed-meshheading:1900915-Neomycin,
pubmed-meshheading:1900915-Open Reading Frames,
pubmed-meshheading:1900915-Promoter Regions, Genetic,
pubmed-meshheading:1900915-Restriction Mapping,
pubmed-meshheading:1900915-Sequence Alignment,
pubmed-meshheading:1900915-Sequence Homology, Nucleic Acid,
pubmed-meshheading:1900915-Streptomyces griseus,
pubmed-meshheading:1900915-Transcription, Genetic,
pubmed-meshheading:1900915-Transformation, Bacterial,
pubmed-meshheading:1900915-alpha-Amylases
|
| pubmed:year |
1991
|
| pubmed:articleTitle |
Cloning, characterization and expression of an alpha-amylase gene from Streptomyces griseus IMRU3570.
|
| pubmed:affiliation |
Area de Microbiología, Facultad de Biología, Universidad de León, Spain.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|