[The effect of a promoter P28-1 of Bacillus subtilis on Streptomyces differentiation].
A 1.1kb promoter P28-1 was inserted into pUC19. After then, the P28-1 was subcloned into the HindIII-EcoRI sites of the high copy number Streptomyces promoter probe plasmid pIJ4083 containing xy1E reporter gene. This recombinant plasmid was designated as pIJ4498. When pIJ4498 was introduced into Streptomyces coelicolor J1501 protoplasts, transformants conferred a white phenotype, whereas the vector pIJ4083 gave rise to colonies of normal, dark grey appearance which is the same as that of J1501 itself. After confirming pIJ4498 structure with some restriction enzymes, it was also introduced into whiG mutant (C71). Crude enzyme extracts were isolated from J1501/pIJ4498, J1501/pIJ4083 and C71/pIJ4498 respectively, the crude enzyme extract from J1501/pIJ4498 could oxidize catechol (colourless) to 2-hydroxy muconic semialdehyde (yellow colour), but crude enzyme extracts from J1501/pIJ4083 and C71/pIJ4498 could not oxidize catechol to 2-hydroxymuconic semialdehyde. The results indicated that P28-1 might be recognised by sigma whiG RNA polymerase, and activated the xylE reporter gene expression and reduced J1501 sporulation.
Tan
H
H
Institute of microbiology, Academia Sinica, Beijing.
Xu
C
C
Tian
Y
Y
Xu
J
J
Chater
K F
KF
D001412:Bacillus subtilis / Q000235:genetics*; D004156:Catechol Oxidase / Q000096:biosynthesis; D015870:Gene Expression; D017930:Genes, Reporter; D010957:Plasmids; D011401:Promoter Regions, Genetic; D013302:Streptomyces / Q000235:genetics / Q000254:growth & development*
D004156:Catechol Oxidase