Mutational cloning in Streptomyces and the isolation of antibiotic production genes

Chater, Keith F.; Bruton, Celia J.

doi:10.1016/0378-1119(83)90037-9

Cited by 132 publications

(50 citation statements)

References 28 publications

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“…Gene Disruption and Deletion-For insertional inactivation, an internal ORF fragment was cloned into the C31 derivative PM1 vector, and the resulting recombinant phage was used to lysogenize the S. coelicolor J1501 strain by insert-directed recombination (66). To obtain the deleted mutant, fragments flanking the region to be deleted were cloned into the PM1 vector, and the recombinant phage was used to lysogenize the wild-type strain by insert-directed recombination.…”

Section: Methodsmentioning

confidence: 99%

A relA/spoT Homologous Gene from Streptomyces coelicolor A3(2) Controls Antibiotic Biosynthetic Genes

Martı́nez-Costa

Arias

Romero

et al. 1996

Journal of Biological Chemistry

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A 0.972-kilobase pair DNA fragment from Streptomyces lividans that induces the production of the bluepigmented antibiotic actinorhodine in S. lividans when cloned on a multicopy plasmid has led to the isolation of a 4-kilobase pair DNA fragment from Streptomyces coelicolor containing homologous sequence. Computer-assisted analysis of the DNA sequence revealed three putative open reading frames (ORFs), ORF1, ORF2, and ORF3. ORF2 extends beyond the sequenced DNA fragment, and its deduced product shares no similarities with any other known proteins in the data bases. ORF3 is also truncated, and its 41-amino acid C-terminal product is identical to the S. coelicolor adenine phosphoribosyltransferase. The 847-amino acid ORF1 protein, with a predicted molecular mass of 94.2 kDa, strongly resembled the relA and spoT gene products from Escherichia coli and the homologs from Vibrio sp. strain S14, Haemophilus influenzae, Streptococcus equisimilis H46A, and Mycoplasma genitalium. Unlike these proteins, the ORF1 amino acid sequence analysis revealed the presence of a putative ATP/GTP-binding domain. A mutant was generated by deleting most of the ORF1 gene that showed an actinorhodine-nonproducing phenotype, while undecylprodigiosin and the calcium-dependent antibiotic were unaffected. The mutant strain grew at a much lower rate than the wild-type strain, and spore formation was delayed. When the gene was propagated on a low copy number vector, not only was actinorhodine production restored, but actinorhodine and undecylprodigiosin production was enhanced in both the mutant and wild-type strains and morphological differentiation returned to wild-type characteristics. (p)ppGpp synthetase activity was not detected in purified ribosomes from the ORF1-deleted mutant, while it was restored by complementation of this strain.

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Section: Methodsmentioning

confidence: 99%

A relA/spoT Homologous Gene from Streptomyces coelicolor A3(2) Controls Antibiotic Biosynthetic Genes

Martı́nez-Costa

Arias

Romero

et al. 1996

Journal of Biological Chemistry

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“…The first example, SLP1, was revealed when pocks were found in a lawn of S. Suarez & Chater (1980), Thompson et al (1980Thompson et al ( ) 1981 First report of amplified DNA segments Robinson et al (1981) 1982 Isolation of promoter sequences Bibb & Cohen (1982 First Streptomyces gene sequenced (aph of S. fradiae) Thompson & Gray (1983) Antibiotic-pathway genes cloned Chater & Bruton (1983), Feitelson & Hopwood (1983), Gil & Hopwood (1983 Complete antibiotic pathway (act) cloned Malpartida & Hopwood (1984)  program for gene recognition Bibb et al (1984) Bibb et al (1985), Buttner et al (1987) Linkage of antibiotic-biosynthetic, -regulatory and -resistance genes Chater & Bruton (1985) John Innes Laboratory Manual published Hopwood et al (1985b) 1987 First Streptomyces transposon discovered Chung (1987) 1987\88 Operon structure (gal and gyl) Fornwald et al (1987), Smith & Chater (1988 Translation without Shine-Dalgarno sequences Janssen et al (1989) Conjugation between E. coli and Streptomyces Mazodier et al (1989) lividans after contact with an S. coelicolor strain devoid of SCP1 and SCP2 (Bibb et al, 1981). The pocks yielded a family of SLP1 plasmids of varying size.…”

Section: Figmentioning

confidence: 99%

“…The first antibioticbiosynthetic genes were cloned, by a variety of methods Chater & Bruton (1985). (Chater & Bruton, 1983 ;Feitelson & Hopwood, 1983 ;Gil & Hopwood, 1983), and soon genes for a whole pathway, the act genes for actinorhodin biosynthesis, were isolated and expressed in a different Streptomyces host (Malpartida & Hopwood, 1984). Just as loss of the blue colour of actinorhodin had been instrumental in the isolation and in vivo mapping of antibiotic-pathway genes (Rudd & Hopwood, 1979), so its reacquisition was crucial to the isolation and in vitro mapping of the genes (Malpartida & Hopwood, 1984, 1986.…”

Section: The In Vitro Yearsmentioning

confidence: 99%

Forty years of genetics with Streptomyces: from in vivo through in vitro to in silico

1999

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“…So far, methylenomycin is the only streptomycete antibiotic the synthesis of which is known to be plasmid encoded. The plasmids involved (pSV1 in S. violaceus-ruber and SCP1 in S. coelicolor) are large and self-transmissible (5,16). SCP1 has recently been physically characterized (20) as a linear plasmid of 350 kb with a copy number of about 4. Both plasmids can be transferred to other streptomycetes when both the resistance determinant and biosynthetic functions are expressed (1,22,28).…”

mentioning

confidence: 99%

An integrated approach to studying regulation of production of the antibiotic methylenomycin by Streptomyces coelicolor A3(2)

Hobbs¹,

Obanye²,

Petty³

et al. 1992

J Bacteriol

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A physiological and molecular biological study was made of the control of methylenomycin biosynthesis by Streptomyces coelicolor A3(2). A simple and reliable assay for this antibiotic was developed. Conditions that permit the synthesis of methylenomycin by S. coelicolor cultures grown in defined medium were elucidated: a readily assimilated carbon and nitrogen source is required. Under these conditions methylenomycin is produced late in the growth phase, at the time of transition from exponential to linear growth. Provided that the phosphate concentration in the medium is kept high, there is synthesis of methylenomycin but not of the other secondary metabolites that this strain can produce. These conditions were used to study the transcription of the methylenomycin gene cluster during the transition from primary to secondary metabolism. The biosynthetic genes of at least one of the mmy transcription units appear to be transcribed before the mmr resistance determinant. The possibility that methylenomycin induces the transcription of mmr is discussed.

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Mutational cloning in Streptomyces and the isolation of antibiotic production genes

Cited by 132 publications

References 28 publications

A relA/spoT Homologous Gene from Streptomyces coelicolor A3(2) Controls Antibiotic Biosynthetic Genes

A relA/spoT Homologous Gene from Streptomyces coelicolor A3(2) Controls Antibiotic Biosynthetic Genes

Forty years of genetics with Streptomyces: from in vivo through in vitro to in silico

An integrated approach to studying regulation of production of the antibiotic methylenomycin by Streptomyces coelicolor A3(2)

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