Expression of the Streptomyces griseus alpha-amylase gene in Escherichia coli.
The amy gene of Streptomyces griseus was not expressed in Escherichia coli cells due to the lack of recognition of the amy promoter by the E. coli RNA polymerase, as confirmed by using promoter-probe vectors. The expression of the amy gene in E. coli was detected only when the promoter-less gene was placed under the control of the lacZ promoter and was dependent on the level of IPTG added to the medium. The extracellular alpha-amylase detected in the culture broth seems to be released by cellular lysis. When the amy gene lacking both leader peptide and promoter was transcribed from the lacZ promoter, no alpha-amylase activity was detected but larger E. coli cells and inclusion bodies were observed.
Vigal
T
T
Area de Microbiología, Facultad de Biología, Universidad de León, Spain.
Martin
J F
JF
Gil
J A
JA
D000595:Amino Acid Sequence; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics*; D007763:Lac Operon; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D013305:Streptomyces griseus / Q000201:enzymology / Q000235:genetics*; D000516:alpha-Amylases / Q000096:biosynthesis* / Q000235:genetics*
D011994:Recombinant Proteins; D000516:alpha-Amylases