Physical and genetical characterisation of a second sex factor, SCP2, for Streptomyces coelicolor A3(2)

Bibb, Mervyn J.; Freeman, R. F.; Hopwood, David A.

doi:10.1007/bf00330831

Cited by 239 publications

(109 citation statements)

References 23 publications

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“…This raises the interesting possibility that an operon may be involved. Recent developments in the study of plasmids in S. coelicolor (Bibb et al, 1977) and of conditions for the efficient transformation of protoplasts by plasmid DNA (Bibb et al, 1978) open the possibility of gene cloning in this organism. Analysis of the physical organization of the actinorhodin biosynthetic genes and of their regulation by in vitro studies are therefore exciting possibilities for the future.…”

Section: B a M R U D D And D A Hopwood Discussionmentioning

confidence: 99%

Genetics of Actinorhodin Biosynthesis by Streptomyces coelicolor A3(2)

Rudd

Hopwood

1979

Journal of General Microbiology

193

117

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35 ~ ~~A series of 76 mutants of Streptomyces coelicolor A3(2) specifically blocked in the synthesis of the binaphthoquinone antibiotic actinorhodin were classified into seven phenotypic classes on the basis of antibiotic activity, accumulation of pigmented precursors or shunt products of actinorhodin biosynthesis, and cosynthesis of actinorhodin in pairwise combinations of mutants. The polarity of cosynthetic reactions, and other phenotypic properties, allowed six of the mutant classes to be arranged in the most probable linear sequence of biosynthetic blocks. One member of each mutant class was mapped unambiguously to the chromosomal linkage map in the short segment between the hisD and guaA loci, suggesting that structural genes for actinorhodin biosynthesis may form an uninterrupted cluster of chromosomal genes.

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Section: B a M R U D D And D A Hopwood Discussionmentioning

confidence: 99%

Genetics of Actinorhodin Biosynthesis by Streptomyces coelicolor A3(2)

Rudd

Hopwood

1979

Journal of General Microbiology

193

117

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“…A critical requirement for this use of R4 is the development of a transfection system. The polyethylene glycol-dependent system used by Bibb et al (1978) to transform Streptomyces protoplasts with SCP2* plasmid DNA has also proved effective in transfection with VP5 DNA (Chater, 19783), and has recently been extended to several other phages including R4 (J. E. Suarez, unpublished results).…”

Section: Discussionmentioning

confidence: 99%

“…We never were able to obtain lysogens of S. coelicolor A3(2): indeed, some derivatives of S. coelicolor A3(2) were very poor hosts for R4. This was independent of the status of the derivatives with respect to plasmids SCPl and SCP2 Bibb et al, 1977). We were unable reliably to score this difference in susceptibility by replica-plating, so that attempts to locate genes controlling it on the S. coelicolor linkage map failed.…”

Section: Host-rangementioning

confidence: 91%

A New, Wide Host-range, Temperate Bacteriophage (R4) of Streptomyces and its Interaction with some Restriction-Modification Systems

Chater

Carter

1979

Journal of General Microbiology

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A new temperate phage, R4, of Streptomyces was isolated from soil on a restrictiondeficient mutant of S. albus G. In its morphology, adsorption properties and growth kinetics R4 resembled other temperate phages of Streptomyces though its requirements for Ca2+ and Mg2+ were higher than usual. It was unable to form plaques above 34.5 "C. R4-mediated transduction was not detected. Unlike other Streptomyces temperate phages, R4 had a wide host-range, which correlated better with the absence of detectable class I1 restriction enzymes than with conventional taxonomic divisions. Many of the sensitive strains [but not, apparently, S. coelicolor A3(2)] could be lysogenized.With the wild-type R4, plaques were obtained on S. albus G only after growth on a restriction-deficient, modification-proficient mutant, and then only at a very low efficiency of plating. All of these plaques were of a mutant type (R4G) which (unlike the parental R4 phage) showed conventional patterns of restriction-modification in the S. albus G (SalGI) and S. albus P (SalPI) systems. R4G mutants, but not R4, were sensitive to a restrictionmodification system present in two S. rimosus strains (2251 and NRRL 2234). DNA from SaZGI-unmodified (but not from modified) R4 or R4G was cleaved by SalGI into more than 30 fragments (mean size 1.35 kilobases; summed molecular weight 3 0 . 0 2~ lo6). R4 DNA was cleaved at one site by EcoRI, at one site by SalPI (= Pst I), and not at all by Hind111 or BamHI.

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“…Most of the media have been described previously; glucose-yeast extract medium (GYM) (Lerch & Ettlinger, 1972); yeast extract-malt extract plus 34% (w/v) sucrose (YEME) (Bibb et al, 1977); R2YE medium (Thompson et al, 1980); P medium (Okanishi et al, 1974) and a modification (MMT) of the minimal medium (MM) described by Hopwood (1967). GYM was employed with D-glucose, 0.4% plus CuS0,.5H20, 0.00025%; MMT (200 ml) for tyrosinase expression was minimal medium with 4 ml Difco Casamino acids (30%, w/v); 1.5 ml of a solution containing 1 % L-histidine; 0.75% each of L-arginine, L-cystine, L-leucine, L-methionine, Lhomoserine, L-phenylalanine, L-proline; 0.1 5% each of adenine and uracil ; 0-01 % each of nicotinamide and thiamin, 10 ml of an L-tyrosine solution (7.5 mg ml-l) and 1.5% Bacto agar.…”

Section: Methodsmentioning

confidence: 99%

“…Total DNA was isolated from mycelium using the procedure described by Chater et al (1982). Large-scale plasmid isolation was by the method of Bibb et al (1977). Plasmid DNA was prepared on a small scale by an alkaline lysis procedure using a modification (Chater et al, 1982) of the method of Birnboim & Doly (1979).…”

Section: Methodsmentioning

confidence: 99%

Cloning and Expression of the Tyrosinase Gene from Streptomyces antibioticus in Streptomyces lividans

1983

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2703 ~In two separate studies a BclI-generated DNA fragment coding for the enzyme tyrosinase, responsible for melanin synthesis, was cloned from Streptomyces antibioticus DNA into two SLPl.2-based plasmid vectors (pIJ37 and pIJ41) to generate the hybrid plasmids, designated pIJ700 and pIJ701, using S. lividam 66 as the host. The fragment (1.55 kb) was subcloned into the multicopy plasmid pIJ350 (which carries thiostrepton resistance and has two non-essential BclI sites) to generate four new plasmids (pIJ702-pIJ705) with the tyrosinase insert located in either orientation at each site. All six plasmids conferred melanin production (the Me]+ phenotype) on their host. As in the S. antibioticus parent, strains of S. lividans carrying the gene specifying tyrosinase synthesis possessed an enzyme activity which was inducible. Most of the tyrosinase activity was secreted during growth of S. antibioticus; in contrast, the majority remained intracellular in the S. lividans clones. The specific activity of the induced tyrosinase activity (intracellular) was higher (up to 36-fold) when the gene was present on the multicopy vector in comparison with its location on the low copy plasmids, pIJ700 or pIJ701, or in S. antibioticus.Restriction mapping of the tyrosinase fragment in pIJ702 revealed endonuclease cleavage sites for several enzymes, including single sites for BgZII, SphI and SstI that are absent from the parent vector (pIJ350). Insertion of DNA fragments at any one of these sites abolished the Mel+ phenotype. The results indicate that pIJ702 is a useful cloning vector with insertional inactivation of the Mel+ character as the basis of clone recognition.

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Physical and genetical characterisation of a second sex factor, SCP2, for Streptomyces coelicolor A3(2)

Cited by 239 publications

References 23 publications

Genetics of Actinorhodin Biosynthesis by Streptomyces coelicolor A3(2)

Genetics of Actinorhodin Biosynthesis by Streptomyces coelicolor A3(2)

A New, Wide Host-range, Temperate Bacteriophage (R4) of Streptomyces and its Interaction with some Restriction-Modification Systems

Cloning and Expression of the Tyrosinase Gene from Streptomyces antibioticus in Streptomyces lividans

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