Restriction enzyme and DNA hybridization analysis of cellulolytic Streptomyces isolates of different origin.
Streptomyces rochei A2 endoglucanase (eglS) and beta-glucosidase (bgs1) genes were used as probes to survey their distribution among 16 Streptomyces strains isolated from different sources and characterized for their cellulolytic activities. The eglS probe hybridized to the genomic DNA of 12 strains with a restriction pattern different from that of S. rochei A2. The DNA from all strains, except one, hybridized with the bgs1 probe and one strain showed the same restriction pattern as seen in S. rochei A2. The sequence localized by the eglS probe in S. thermoviolaceus and the one localized by the bgs1 probe in strain EC1 were cloned and expressed in E. coli in plasmids pTAE and pCSF203, respectively. The restriction maps showed that the cloned genes were identical to eglS and bgs1. The restriction enzyme analysis and genomic DNA from all the strains identified nine different groups, each characterized by a distinctive pattern and in agreement with the results of the hybridization experiments.
Marri
L
L
Dipartimento di Biologia Molecolare, Università di Siena, Italy.
Barboni
E
E
Irdani
T
T
Perito
B
B
Mastromei
G
G
D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D016680:Genome, Bacterial; D015183:Restriction Mapping; D013302:Streptomyces / Q000235:genetics* / Q000302:isolation & purification
D004269:DNA, Bacterial