Conjugal transfer using the bacteriophage ϕC31 att/int system and properties of the attB site in Streptomyces ambofaciens

2007 ◽  
Vol 30 (4) ◽  
pp. 695-699 ◽  
Author(s):  
Mi-Kyung Kim ◽  
Heon-Su Ha ◽  
Sun-Uk Choi
Biologia ◽  
2014 ◽  
Vol 69 (8) ◽  
Author(s):  
Jae-Young Park ◽  
Sun-Uk Choi

AbstractAn effective transformation method for molecular genetic studies of Streptomyces cinnamoneus producing transglutaminase was established using intergeneric conjugal transfer based on the bacteriophage ϕC31 att/int system. The high efficiency of S. cinnamoneus transconjugation was obtained on mannitol soya flour medium containing 50 mM MgCl2, using 2.5 × 108 Escherichia coli as donor and spores treated with heat treatment at 35°C for 10 min as host. By cloning and sequencing the attB integration site, a single attB site within an open reading frame coding for a pirin homologue was located in S. cinnamoneus genome. Its sequence exhibited the highest degree of sequence similarity with that of Streptomyces clavuligerus. These results provide sufficient efficiency to enable conjugal transfer of genetic elements for S. cinnamoneus, and also should facilitate molecular genetic studies for improvement in production ability of transglutaminase used in food industry.


1991 ◽  
Vol 17 (1) ◽  
pp. 69-70 ◽  
Author(s):  
M.S. Dryden ◽  
M. McCann ◽  
I. Phillips
Keyword(s):  

1997 ◽  
Vol 43 (12) ◽  
pp. 1118-1125 ◽  
Author(s):  
Martine Aubert ◽  
Elisabeth Weber ◽  
Brigitte Gintz ◽  
Bernard Decaris ◽  
Keith F. Chater

The deduced product of the spa2 gene of Streptomyces ambofaciens is a homologue of RspA, involved in stationary-phase σs factor regulation in Escherichia coli. This suggests that Spa2 could play a part in stationary-phase-associated differentiation in S. ambofaciens. The disruption of spa2 led to reductions in aerial mycelial development and associated spore pigmentation. The mutant phenotype reverted to the wild-type phenotype when the disruption construct spontaneously excised. The spa2 disruption had no detectable effect on growth rates in different media or antibiotic production and resistance. When spa2 was placed on a multicopy plasmid, a severe defect in formation and pigmentation of aerial mycelium resulted. These results strongly suggest that Spa2 is involved in a complex manner in the morphological differentiation process.Key words: Streptomyces, differentiation, stationary-phase regulator.


1998 ◽  
Vol 28 (3) ◽  
pp. 571-582 ◽  
Author(s):  
Linda M. Parsons ◽  
Craig S. Jankowski ◽  
Keith M. Derbyshire

Microbiology ◽  
2010 ◽  
Vol 156 (9) ◽  
pp. 2723-2733 ◽  
Author(s):  
Jing Wang ◽  
Gregg S. Pettis

Conjugal transfer of circular plasmids in Streptomyces involves a unique mechanism employing few plasmid-encoded loci and the transfer of double-stranded DNA by an as yet uncharacterized intercellular route. Efficient transfer of the circular streptomycete plasmid pIJ101 requires only two plasmid loci: the pIJ101 tra gene, and as a cis-acting function known as clt. Here, we compared the ability of the pIJ101 transfer apparatus to promote conjugal transfer of circular versus linear versions of the same replicon. While the pIJ101 tra locus readily transferred the circular form of the replicon, the linear version was transferred orders of magnitude less efficiently and all plasmids isolated from the transconjugants were circular, regardless of their original configuration in the donor. Additionally, relatively rare circularization of linear plasmids was detectable in the donor cells, which is consistent with the notion that this event was a prerequisite for transfer by TraB(pIJ101). Linear versions of this same replicon did transfer efficiently, in that configuration, from strains containing the conjugative linear plasmid SLP2. Our data indicate that functions necessary and sufficient for transfer of circular DNA were insufficient for transfer of a related linear DNA molecule. The results here suggest that the conjugation mechanisms of linear versus circular DNA in Streptomyces spp. are inherently different and/or that efficient transfer of linear DNA requires additional components.


Gene ◽  
1987 ◽  
Vol 59 (1) ◽  
pp. 137-144 ◽  
Author(s):  
Jean-Marc Simonet ◽  
Frédéric Boccard ◽  
Jean-Luc Pernodet ◽  
Josette Gagnat ◽  
Michel Guérineau

2010 ◽  
Vol 105 (2) ◽  
pp. 171-175 ◽  
Author(s):  
Clelton A. Santos ◽  
Gislayne T. Vilas-Bôas ◽  
Didier Lereclus ◽  
Marise T. Suzuki ◽  
Elisangela A. Angelo ◽  
...  

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