Cloning of the replication origin (oriC) of Streptoverticillum caespitosus and transformation of Streptomyces lividans ZX7 / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 662-666, 2002.
Artigo
em Chinês
| WPRIM
| ID: wpr-256142
ABSTRACT
Streptoverticillum caespitosus ATCC27422 is a producing strain of mitomycin A for cancer therapy. Taking the advantage of the conserved sequences of genes flanking the oriC of high G + C Gram-positive bacteria, a 1.3 kb DNA fragment containing oriC and its flanking region was cloned by PCR. Nuleotide sequence comparisons revealed that the cloned fragment is more than 80% identical to the same region of S. coelicolor. There are 22 DnaA-boxes in the oriC region, and the conserved sequence of DnaA-box is TTGTCCACA. The plasmid containing the oriC of S. caespitosus was constructed (pMJ9), and it was able to transform the protoplast of Streptomyces lividans ZX7 at the frequency of 3.2 x 10(2) transformants/micrograms plasmid DNA. The colony and mycelia's morphology of the transformants are normal. The constructed plasmid can exist stable in the host as a low copy extra-chromosome replicon. The high rate of the homology and the cross genus replication initiation activity suggests close relationship between Streptomyces and Streptoverticillum in the evolution. While the maximum likelihood phylogenetic tree based upon the oriC of S. caespitosus and several Streptomyces spp. revealed that S. caespitosus differed extensively from the Streptomyces spp. This result supports the separation of Streptoverticillum from Streptomyces.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Streptomyces
/
Transformação Bacteriana
/
Actinomycetales
/
Dados de Sequência Molecular
/
Sequência de Bases
/
Southern Blotting
/
Clonagem Molecular
/
Sequência Conservada
/
Origem de Replicação
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2002
Tipo de documento:
Artigo
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