Expression of Streptomyces genes encoding extracellular enzymes in Brevibacterium lactofermentum: secretion proceeds by removal of the same leader peptide as in Streptomyces lividans.
The alpha-amylase gene (amy) from Streptomyces griseus IMRU 3570 and the beta-galactosidase gene (lac) from S. lividans were subcloned into Brevibacterium lactofermentum or B. lactofermentum/Escherichia coli shuttle vectors. The amy gene was not expressed in B. lactofermentum from its own promoter but was efficiently expressed when the promoter of the kanamycin resistance gene (kan) was inserted upstream of the promoterless amylase gene. The lac gene from S. lividans was subcloned without its native promoter and was expressed when placed downstream of pBL1 promoters P2 or P3. The alpha-amylase was secreted extracellularly by removal of the same 28-amino acid leader peptide as in S. lividans. The amy and lac genes provide useful markers for selection of transformants and will facilitate the study of protein secretion in B. lactofermentum.
Area de Microbiología, Facultad de Biología, Universidad de León, Spain.
R F
RF
Cadenas
J A
JA
Gil
J F
JF
Martín
D000595:Amino Acid Sequence; D001709:Biotechnology; D001951:Brevibacterium / Q000235:genetics*; D003001:Cloning, Molecular; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors; D007613:Kanamycin Resistance / Q000235:genetics; D007763:Lac Operon; D008969:Molecular Sequence Data; D010957:Plasmids; D021382:Protein Sorting Signals / Q000235:genetics; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*; D013305:Streptomyces griseus / Q000201:enzymology / Q000235:genetics; D000516:alpha-Amylases / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D021382:Protein Sorting Signals; D000516:alpha-Amylases; D001616:beta-Galactosidase