The alpha-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 in a loss of amylolytic activity in the host cells that can be assayed using a simple and inexpensive staining procedure.
Ikuta
N
N
Genetics Institute, Escola Superior de Agricultura Luiz de Queiroz, Universidade de Sao Paulo, Brazil.
Souza
M B
MB
Valencia
F F
FF
Castro
M E
ME
Schenberg
A C
AC
Pizzirani-Kleiner
A
A
Astolfi-Filho
S
S
D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics*; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000032:analysis*; D004926:Escherichia coli / Q000235:genetics*; D005819:Genetic Markers; D005822:Genetic Vectors; D010957:Plasmids; D014169:Transformation, Bacterial; D000516:alpha-Amylases / Q000235:genetics* / Q000378:metabolism
D004274:DNA, Recombinant; D005819:Genetic Markers; D000516:alpha-Amylases