Cloning and expression of the sco2127 gene from Streptomyces coelicolor M145.
It is known that Streptomyces peucetius var. caesius mutants resistant to 2-deoxyglucose (Dog(R)) exhibit glucose transport deficiency, low glucose kinase (Glk) activity and insensitivity to carbon catabolite repression (CCR). This phenotype can be pleiotropically complemented by a 576-bp gene encoding SCO2127 from Streptomyces coelicolor, suggesting the participation of this protein in the CCR process. In the present work, the sco2127 region was subcloned into pQE30 and its transcription product (SCO2127-His(6)) overexpressed. This procedure allowed purification of SCO2127 (with a Ni-sepharose resin) and production of polyclonal antibodies. In western blot assays, the antibodies gave a positive reaction against protein extracts from both S. coelicolor and S. peucetius var. caesius, appearing as a single band of 34 kDa. No protein was detected using extracts from a S. coelicolor mutant lacking the sco2127 gene (Deltasco2127). In agreement with its possible involvement in the CCR process, SCO2127 was detected during the logarithmic growth phase of S. coelicolor grown in minimal medium supplemented with 50 and 100 mM glucose. In addition, when 50 mM glucose was utilized, SCO2127 and residual glucose concentration simultaneously decreased at later stages of the microbial growth.
Chávez
Adán
A
Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Mexico DF, Mexico.
García-Huante
Yolanda
Y
Ruiz
Beatriz
B
Langley
Elizabeth
E
Rodríguez-Sanoja
Romina
R
Sanchez
Sergio
S
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial; D005947:Glucose / Q000378:metabolism; D008970:Molecular Weight; D048372:Streptomyces coelicolor / Q000737:chemistry / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
D001426:Bacterial Proteins; D005947:Glucose