The α-Amylase Gene as a Marker for Gene Cloning: Direct Screening of Recombinant Clones

We report the construction and use of a new system for the direct screening of recombinant clones after transformation. The system uses a Bacillus subtilis-Escherichia coli shuttle vector that carries the B. subtilis structural gene for alpha-amylase. Insertion of foreign DNA into this gene results...

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Bibliographic Details
Published in:	Bio/technology (New York, N.Y. 1983) Vol. 8; no. 3; pp. 241 - 242
Main Authors:	Ikuta, Nilo, Souza, Maria Beatriz N, Valencia, Fernando F, Castro, Maria Elita B, Schenberg, Ana Clara G, Pizzirani-Kleiner, Aline, Astolfi-Filho, Spartaco
Format:	Journal Article
Language:	English
Published:	New York, NY Nature Publications 01-03-1990
Subjects:	alpha-Amylases - genetics alpha-Amylases - metabolism Bacillus subtilis - enzymology Bacillus subtilis - genetics Biological and medical sciences Biotechnology Chromosome Mapping Cloning, Molecular DNA, Recombinant - analysis Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Genetic engineering Genetic Markers Genetic technics Genetic Vectors Methods. Procedures. Technologies Plasmids Transformation, Bacterial Plasmid Screening Genetic transformation Gene Molecular probe Recombinant DNA α-Amylase Molecular cloning
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Summary:	We report the construction and use of a new system for the direct screening of recombinant clones after transformation. The system uses a Bacillus subtilis-Escherichia coli shuttle vector that carries the B. subtilis structural gene for alpha-amylase. Insertion of foreign DNA into this gene results in a loss of amylolytic activity in the host cells that can be assayed using a simple and inexpensive staining procedure.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0733-222X 2331-3684
DOI:	10.1038/nbt0390-241