WO2006022476A1 - Transcriptional activator derived from streptomyces peucetius and the gene thereof - Google Patents

Transcriptional activator derived from streptomyces peucetius and the gene thereof Download PDF

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WO2006022476A1
WO2006022476A1 PCT/KR2005/001800 KR2005001800W WO2006022476A1 WO 2006022476 A1 WO2006022476 A1 WO 2006022476A1 KR 2005001800 W KR2005001800 W KR 2005001800W WO 2006022476 A1 WO2006022476 A1 WO 2006022476A1
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afsr
gene
streptomyces
transcriptional activator
present
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PCT/KR2005/001800
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French (fr)
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Jin Suk Woo
Jae Kyung Sohng
Hei Chan Lee
Kwang Kyoung Liou
Young Soo Jung
Sun Youp Kang
Dae Hee Kim
Ji Young Yang
Wun Min Seo
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Genechem Inc.
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Publication of WO2006022476A1 publication Critical patent/WO2006022476A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/36—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Actinomyces; from Streptomyces (G)

Definitions

  • the present invention relates to a transcriptional activator derived from Streptomyces and the gene thereof (afsR-p), more particularly, to afsR-p gene derived from Streptomyces peucetius having the function of a transcriptional activator to regulate the formation of antibiotics.
  • Streptomyces are filamentous soil bacteria that produce a wide variety of secondary metabolites (Crandall, L. W. and Hamill, R.L., Antibiotics produced by Streptomyces: major structural classes, In S. W. Queener and L.E. Day (ed.), The Bacteria 9, 355-402, 1986). Little is known about the regulation in Streptomyces. Generally, the regulatory genes involved in secondary metabolites production are divided into two classes, pathway-specific and global (Chater, K.R and Bibb, MJ., Bio/Technology, 7:57, 1996).
  • Pathway specific regulatory genes are operated by two-component regulatory systems, which target aspartate residues for modification. Recent studies have shown that serine/threonine and tyrosine kinases are present in both prokaryotes and eukaryotes (Cozzone, AJ., Folia Microbiol, 42:165, 1997).
  • AfsR is one of the substrates of AfsK, which constitutes the AfsK-AfsR system. It is the first example of a functionally relevant serine/threonine phosphorylation system in prokaryotes (Kennelly, PJ. and Potts, M., J. BacterioL, 178:4759, 1996). Subsequent studies have shown that AfsK-AfsR system is widely used in Streptomyces and influences secondary metabolism and morphogenesis. Furthermore, it has been revealed that AfsR serves as a transcriptional activator for afsS, which activates the pathway-specific genes, act ⁇ l-ORF4, in the actinorhodin biosynthetic gene cluster (Lee, P.C. et al., MoI. Microbiol., 43:1413, 2002).
  • ⁇ -actinorhodin is antibiotics, which are used in preparation of medicine. Most antibiotics are not proteins and are synthesized through complex process where many enzymes, which is gene products different from each other, participate
  • AfsR from S. coelicolor A3 encodes 993 amino acids. Unlike other DNA-binding proteins, AfsR has conserved two ATP-binding motifs and a catalytic domain. AfsR is therefore called a global regulatory gene for secondary metabolite production.
  • the NH 2 -terminal portion of AfsR shows similarity to the pathway-specific regulatory genes of Streptomyces, such as ⁇ cflI-ORF4, redD, dnrl and ccaR (Fernandez-Moreno, M. A. et al, Cell, 66:769, 1991; Narva, K.E. and Feitelson, J.S., J.
  • the present inventors have cloned afsR gene from S. peucetius, revealed conserved sequences between AfsRs and transformed S.lividans TK24 with the cloned gene, followed by culturing and thus confirmed that the production of ⁇ -actinorhodin increased, thereby completing the present invention.
  • An object of the present invention is to provide a transcriptional activator, AfsR-p which regulates the formation of antibiotics and the gene thereof.
  • Another object of the present invention is to provide a recombinant vector containing the gene and a microorganism transformed with the recombinant vector.
  • Still another object of the present invention is to provide primers for indentifying afsR.
  • the present invention provides AfsR-p with an amino acid sequence of SEQ ID NO: 2 which has the function of a transcriptional activator regulating the formation of antibiotics.
  • the present invention provides afsR-p gene having a DNA sequence of SEQ ID NO: 1, a recombinant vector containing the gene and a microorganism transformed by introducing the recombinant vector to a host cell selected from the group consisting of bacteria, mold, and yeast.
  • the gene is derived from Streptomyces peucetius and the host cell is the genus Streptomyces having the ability of antibiotic production.
  • the present invention also provides primer set for identifying afsR gene, which has a DNA sequence of SEQ ID NOs: 5 and 6.
  • FIG. 1 shows the multiple alignments comparing homology of AfsR-p derived from Streptomyces peucetius ATCC 27952, AfsR-g derived from Streptomyces griseus and AfsR derived from Streptomyces coelicolor A3.
  • FIG 2 is a graph showing an enhancement of ⁇ -actinorhodin production in Streptomyces lividans TK24 transformed with afsR-p according to the present invention.
  • 1 and 3 are Streptomyces lividans TK24 transformed with pIBR25 and 2 and 4 are Streptomyces lividans TK24 transformed with pGIBR.
  • 1 and 2 are result of culture at 28°C for 4 days and 3 and 4 are result of culture at 28°C for 8 days.
  • Streptomyces peucetius ATCC 27952 is well-known for doxorubicin producer.
  • the inventors found transcriptional activator g ⁇ &as(afsR-p) participating in the production of doxorubicin from Streptomyces peucetius ATCC 27952 through homology analysis with regulatory genes from other genus Streptomyces and transformed Streptomyces lividans producing ⁇ -actinorhodin with the recombinant vector containing the gene to reveal the function of the gene, and then examined whether or not the ⁇ -actinorhodin production in the transformed Streptomyces lividans increased.
  • DNA fragments of approximately 2000 to 3000-bp in length were subcloned into pGEM-3Zf(+) or pGEM7-7Zf(+) (Promega, USA) and sequencing was performed by the didoxynucleotide chain termination method.
  • Genome databases of S. peucetius were formulated by the present inventors for the first time (Parajuli N. et al, Arch. Biochem. Biophys., 425:233, 2004).
  • the open reading frames (ORFs) of regulatory genes obtained using Glimmer 2.0 were annotated by BLAST (Delcher A.L., Nucleic Acids Res., 27:4636, 1999).
  • BLAST Carlcher A.L., Nucleic Acids Res., 27:4636, 1999.
  • Global regulatory gene was initially searched based on two ATP binding domain signatures, GIGGVGKT(A-type) and LLDNA(B-type) and a catalytic domain, AEPET.
  • the genes including such motifs or domain were searched by BLAST of the GenBank and each gene was translated to amino acid using GeneDoc program
  • AfsR-p shares 60% sequence identity with AfsR protein (993 amino acids) from S. coelicolor A3 (2) and 61% with AfsR-g (979 amino acids) from S. griseus (FIG 1).
  • the afsR-p with ATG start codon and TGA stop codon is oriented in forward direction and there is a distinct ribosome-binding site before start codon.
  • the NH 2 -terminal portion of AfsR-p shows similarity to dnrl. It indicates that AfsR-p is a DNA-binding protein.
  • afsR from S. coelicolor A3(2) is clustered with afsK and other regulatory genes. However, afsR-p is located with hypothetical proteins in S. peucetius.
  • Primers of SEQ ID NOs: 3 and 4 with HindUI/EcoRI sites were used to amplify the afsR-p from genomic DNA of S. peucetius.
  • the PCR conditions were as follows: denaturation at 94 0 C, annealing at 65 0 C and polymerization at 74 0 C and the DNA amplification was performed in a total volume of 50 ⁇ l with 2.5 units of Taq-DNA polymerase, 10% dimethylsulfoxide, 2.5 pmol of primers, 0.2 mM dNTP and 0.1 ⁇ g genomic DNA in appropriate buffer.
  • the amplified afsR-p was cloned to pIBR25 (Sthapit B. et al, FEBS Lett., 566:201, 2004) at EcoRI and HindIII sites to produce pGIBR.
  • pGIBR25 Sthapit B. et al, FEBS Lett., 566:201, 2004
  • the protoplasts of S. lividans TK24 were transformed with pGIBR and pIBR25 to generate S. lividans/GTBR and S. Hvidans/IBR.
  • Two oligonucleotides of SEQ ID NO: 5 and 6 were synthesized based on the conserved residues of afsR-p, afsR-g and afsR as shown in FIG 1 and used to amplify its homologous regions from genomic DNA of Streptomyces achromogens var rubradiris NRRL 3061 (rubradirin producer) and Streptomyces clavuligerous NRRL 3585 (clavulanic acid producer).
  • the PCR conditions were the same as those in Example 3.
  • S represents G or C.
  • the amplified products were cloned into pGEMT-easy vector (Promega, USA).
  • the cloned AfsR-a (171 amino acids) from S. achromogens showed 66% identity with AfsR from S. coelicolor A3(2) and the cloned AfsR-c (178 amino acids) from S. clavuligerous showed 86% identity with AfsR-g from S. griseus.
  • results indicate the successful amplification of afsR specific nucleotide sequences from the genus Streptomyces, which will be useful for cloning of afsR homologue gene from respective genera in future, and the primers of SEQ ID NOs: 5 and 6 reported can be used to isolate afsR from other actinomycetes.
  • Example 5 Effects of afsR-p on ⁇ -actinorhodin production in S. lividans TK24
  • afsR-p gene according to the present invention functioned as a transcriptional activator to regulate the production of doxorubicin in Streptomyces peucetius.
  • the present invention has the effect to provide afsR-p gene having the function of a transcriptional activator to regulate the formation of antibiotics in the genus Strepetomyces.
  • afsR-p gene according to the present invention is useful for increasing the formation of antibiotics in the genus Strepetomyces.

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Abstract

The present invention relates to transcriptional activator derived from Streptomyces and the gene thereof (afsR-p), more particularly, to afsR-p gene derived from Streptomyces peucetius having the function of a transcriptional activator to regulate the formation of antibiotics. The afsR-p gene according to the present invention is useful for increasing the formation of antibiotics, such as Ϝ-actinorhodin, doxorubicin etc. from the genus Strepetomyces.

Description

TRANSCRIPTIONAL ACTIVATOR DERIVED FROM STREPTOMYCES PEUCETIUS AND THE GENE THEREOF
TECHNICAL FIELD
The present invention relates to a transcriptional activator derived from Streptomyces and the gene thereof (afsR-p), more particularly, to afsR-p gene derived from Streptomyces peucetius having the function of a transcriptional activator to regulate the formation of antibiotics.
BACKGROUND ART
Streptomyces are filamentous soil bacteria that produce a wide variety of secondary metabolites (Crandall, L. W. and Hamill, R.L., Antibiotics produced by Streptomyces: major structural classes, In S. W. Queener and L.E. Day (ed.), The Bacteria 9, 355-402, 1986). Little is known about the regulation in Streptomyces. Generally, the regulatory genes involved in secondary metabolites production are divided into two classes, pathway-specific and global (Chater, K.R and Bibb, MJ., Bio/Technology, 7:57, 1996). While almost all pathway-specific regulatory genes located within a biosynthetic gene cluster control only one kind of antibiotic biosynthesis, the global regulatory genes located outside a biosynthetic gene cluster seem to control the secondary metabolism, morphological and physiological differentiation in Streptomyces (Umeyama, T. et al., Appl. Microbiol. BiotechnoL, 59:419, 2002 ).
Pathway specific regulatory genes are operated by two-component regulatory systems, which target aspartate residues for modification. Recent studies have shown that serine/threonine and tyrosine kinases are present in both prokaryotes and eukaryotes (Cozzone, AJ., Folia Microbiol, 42:165, 1997).
In Streptomyces coelicolor A3, more than 40 eukaryotic type serine/threonine and tyrosine kinases are found (http://www.sanger.ac.uk/Projects/S_coelicolor/). The influence of such protein kinases in secondary metabolism was first studied on AfsK.
AfsR is one of the substrates of AfsK, which constitutes the AfsK-AfsR system. It is the first example of a functionally relevant serine/threonine phosphorylation system in prokaryotes (Kennelly, PJ. and Potts, M., J. BacterioL, 178:4759, 1996). Subsequent studies have shown that AfsK-AfsR system is widely used in Streptomyces and influences secondary metabolism and morphogenesis. Furthermore, it has been revealed that AfsR serves as a transcriptional activator for afsS, which activates the pathway-specific genes, actϊl-ORF4, in the actinorhodin biosynthetic gene cluster (Lee, P.C. et al., MoI. Microbiol., 43:1413, 2002).
γ-actinorhodin is antibiotics, which are used in preparation of medicine. Most antibiotics are not proteins and are synthesized through complex process where many enzymes, which is gene products different from each other, participate
AfsR from S. coelicolor A3 encodes 993 amino acids. Unlike other DNA-binding proteins, AfsR has conserved two ATP-binding motifs and a catalytic domain. AfsR is therefore called a global regulatory gene for secondary metabolite production. The NH2-terminal portion of AfsR shows similarity to the pathway-specific regulatory genes of Streptomyces, such as αcflI-ORF4, redD, dnrl and ccaR (Fernandez-Moreno, M. A. et al, Cell, 66:769, 1991; Narva, K.E. and Feitelson, J.S., J. BacterioL, 172:326, 1990; Stutzman-Engwall, KJ. et al, J. Bacterial, 174:144, 1992; Perez-Llarena, FJ. et al, J. BacterioL, 179:2053, 1997). While partial sequences of gene encoding AfsR from the genus Streptomyces are revealed, conserved sequences of that are not reported yet.
Accordingly, the present inventors have cloned afsR gene from S. peucetius, revealed conserved sequences between AfsRs and transformed S.lividans TK24 with the cloned gene, followed by culturing and thus confirmed that the production of γ-actinorhodin increased, thereby completing the present invention.
DISCLOSURE OF THE INVENTION
An object of the present invention is to provide a transcriptional activator, AfsR-p which regulates the formation of antibiotics and the gene thereof.
Another object of the present invention is to provide a recombinant vector containing the gene and a microorganism transformed with the recombinant vector.
Still another object of the present invention is to provide primers for indentifying afsR.
To achieve the above objects, the present invention provides AfsR-p with an amino acid sequence of SEQ ID NO: 2 which has the function of a transcriptional activator regulating the formation of antibiotics.
In another aspect, the present invention provides afsR-p gene having a DNA sequence of SEQ ID NO: 1, a recombinant vector containing the gene and a microorganism transformed by introducing the recombinant vector to a host cell selected from the group consisting of bacteria, mold, and yeast.
In the present invention, the gene is derived from Streptomyces peucetius and the host cell is the genus Streptomyces having the ability of antibiotic production.
In another aspect, the present invention also provides primer set for identifying afsR gene, which has a DNA sequence of SEQ ID NOs: 5 and 6.
BRIEF DESCRIPTION OF DRAWINGS
FIG. 1 shows the multiple alignments comparing homology of AfsR-p derived from Streptomyces peucetius ATCC 27952, AfsR-g derived from Streptomyces griseus and AfsR derived from Streptomyces coelicolor A3.
FIG 2 is a graph showing an enhancement of γ-actinorhodin production in Streptomyces lividans TK24 transformed with afsR-p according to the present invention. 1 and 3 are Streptomyces lividans TK24 transformed with pIBR25 and 2 and 4 are Streptomyces lividans TK24 transformed with pGIBR. 1 and 2 are result of culture at 28°C for 4 days and 3 and 4 are result of culture at 28°C for 8 days.
DETAILED DESCRIPTION OF THE INVENTION
Streptomyces peucetius ATCC 27952 is well-known for doxorubicin producer. In the present invention, the inventors found transcriptional activator g < &as(afsR-p) participating in the production of doxorubicin from Streptomyces peucetius ATCC 27952 through homology analysis with regulatory genes from other genus Streptomyces and transformed Streptomyces lividans producing γ-actinorhodin with the recombinant vector containing the gene to reveal the function of the gene, and then examined whether or not the γ-actinorhodin production in the transformed Streptomyces lividans increased.
The present invention will hereinafter be described in further detail by examples. It will, however, be obvious to a person skilled in the art that these examples are given for illustrative purpose only, and the scope of the present invention is not limited to or by these examples.
Example 1: The construction of cosmid library
The total genome of S. peucetius has been sequenced using various cosmids and the shotgun approach (Himmelreich, H., et al., Nucleic Acids Res., 24:4420, 1996). The all non-redundant fragments were then assembled in the computer using programs: PRED(Ewing B. and Green P., Genome Res., 8:186, 1998) and PHRAP (http://www.phrap.org) to generate contigs. An additional genome library was also prepared to assist the whole genomic sequencing. DNA fragments of approximately 2000 to 3000-bp in length were subcloned into pGEM-3Zf(+) or pGEM7-7Zf(+) (Promega, USA) and sequencing was performed by the didoxynucleotide chain termination method.
Example 2: Identification and analysis of regulatory genes
Genome databases of S. peucetius were formulated by the present inventors for the first time (Parajuli N. et al, Arch. Biochem. Biophys., 425:233, 2004). The open reading frames (ORFs) of regulatory genes obtained using Glimmer 2.0 were annotated by BLAST (Delcher A.L., Nucleic Acids Res., 27:4636, 1999). Global regulatory gene was initially searched based on two ATP binding domain signatures, GIGGVGKT(A-type) and LLDNA(B-type) and a catalytic domain, AEPET. The genes including such motifs or domain were searched by BLAST of the GenBank and each gene was translated to amino acid using GeneDoc program
(Nicholas K.B. and Nicholas H.B., GeneDoc: a tool for editing and annotating multiple sequence alignments, Editor and Shading Utility Version 2.6.002, 1997).
The deduced amino acid sequences of the putative S. peucetius global regulatory genes were aligned with the afsR from S. coelicolor A3(2) and afsR-g from Streptomyces griseus by ClustalX (Thompson J.D. et ah, Nucleic Acids Res., 24:4876, 1997) and then compared with them(FIG 1).
The genome analysis of S. peucetius revealed afsR homologies in five various regions of chromosome, and only afsR-p encoding 983 amino acids has conserved two ATP-binding sites (A-type: 321GIGGVGKT328 and B-type: 399MVLLD403) and a catalytic domain (399IvIVLLD403) unlike other DNA-binding proteins NtrC
{Salmonella typhimurium), NifA {Klebsiella pneumoniae), DnaA {Bacillus subtilis),
FtsH {Pseudomonas multocidd), AtpD and MaIK {E. coli), and RecA {Mycobacterium tuberculosis).
AfsR-p shares 60% sequence identity with AfsR protein (993 amino acids) from S. coelicolor A3 (2) and 61% with AfsR-g (979 amino acids) from S. griseus (FIG 1). The afsR-p with ATG start codon and TGA stop codon is oriented in forward direction and there is a distinct ribosome-binding site before start codon. Furthermore, the NH2-terminal portion of AfsR-p shows similarity to dnrl. It indicates that AfsR-p is a DNA-binding protein.
afsR from S. coelicolor A3(2) is clustered with afsK and other regulatory genes. However, afsR-p is located with hypothetical proteins in S. peucetius.
Example 3: Cloning and expression of afsR from S. peucetius
Primers of SEQ ID NOs: 3 and 4 with HindUI/EcoRI sites were used to amplify the afsR-p from genomic DNA of S. peucetius. The PCR conditions were as follows: denaturation at 94 0C, annealing at 65 0C and polymerization at 74 0C and the DNA amplification was performed in a total volume of 50 μl with 2.5 units of Taq-DNA polymerase, 10% dimethylsulfoxide, 2.5 pmol of primers, 0.2 mM dNTP and 0.1 μg genomic DNA in appropriate buffer. SEQ ID NO: 3: S'-TCCGGAATTCCCGGCAGGGGGC-S' SEQ ID NO: 4: 5'-CGAAGCTTCGGACCGAGCACGA-S'
The amplified afsR-p was cloned to pIBR25 (Sthapit B. et al, FEBS Lett., 566:201, 2004) at EcoRI and HindIII sites to produce pGIBR. Finally, the protoplasts of S. lividans TK24 were transformed with pGIBR and pIBR25 to generate S. lividans/GTBR and S. Hvidans/IBR.
Example 4: Cloning of the conserved regions of afsR from other S. yeuc etius
Two oligonucleotides of SEQ ID NO: 5 and 6 were synthesized based on the conserved residues of afsR-p, afsR-g and afsR as shown in FIG 1 and used to amplify its homologous regions from genomic DNA of Streptomyces achromogens var rubradiris NRRL 3061 (rubradirin producer) and Streptomyces clavuligerous NRRL 3585 (clavulanic acid producer). The PCR conditions were the same as those in Example 3.
SEQ ID NO: 5: 5'-GCSGGSATCGGSGGSGTSGGSAAGACSA-S' SEQ ID NO: 6: 5'-SGCSAGSGCSGASACSGCCATSAC-S'
Herein, S represents G or C.
The amplified products were cloned into pGEMT-easy vector (Promega, USA). The cloned AfsR-a (171 amino acids) from S. achromogens showed 66% identity with AfsR from S. coelicolor A3(2) and the cloned AfsR-c (178 amino acids) from S. clavuligerous showed 86% identity with AfsR-g from S. griseus. The results indicate the successful amplification of afsR specific nucleotide sequences from the genus Streptomyces, which will be useful for cloning of afsR homologue gene from respective genera in future, and the primers of SEQ ID NOs: 5 and 6 reported can be used to isolate afsR from other actinomycetes.
Futhermore, the DNA sequence oϊafsR-p, afsR-a and afsR-c revealed in the present invention have been deposited in the EMBL under the Accession No. AJ786384, AJ786385, AJ786386, respectively.
Example 5: Effects of afsR-p on γ-actinorhodin production in S. lividans TK24
S. Hvidans/GΪBR and S. lividans/ΪBR collected from example 3 were cultured in 50 ml TSB medium supplemented with 50 μg ml"1 thiostrepton at 28 0C and 250 rpm for 2 days. 1 ml of cells was then transferred into 100 ml R2YE medium (US 5,843,735) supplemented with 100 μg ml"1 thiostrepton and incubated at 28 0C at 250 rpm. 5 ml samples were withdrawn every 2 days and the mycelia were removed by centrifugation. Then, the supernatants were acidified with IN HCl to pH 2 and extracted with half volume of methanol: CHCI3 (1:1). The chloroform fraction was collected and was measured at A542 (ε542= 18600 for pure compound) using UV-spectrophotometer (Shimadzu, USA).
Consequently, a threefold increase in production of γ-actinorhodin in S. lividans/GΪBR was determined comparing to control (S. HvidansΔBR) as shown in FIG 2. This result indicated that afsR-p gene according to the present invention functioned as a transcriptional activator to increase the production of γ-actinorhodin in S. lividans.
This result also indicated that that afsR-p gene according to the present invention functioned as a transcriptional activator to regulate the production of doxorubicin in Streptomyces peucetius.
Although the present invention has been described in detail with reference to the specific features, it will be apparent to those skilled in the art that this description is only for a preferred embodiment and does not limit the scope of the present invention. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.
INDUSTRIAL APPLICABILITY
As described above in detail, the present invention has the effect to provide afsR-p gene having the function of a transcriptional activator to regulate the formation of antibiotics in the genus Strepetomyces. afsR-p gene according to the present invention is useful for increasing the formation of antibiotics in the genus Strepetomyces.

Claims

THE CLAIMS
What is claimed is:
L A transcriptional activator, AfsR-p, having an amino acid sequence of SEQ ID NO: 2.
2. The AfsR-p according to claim 1, which has the function of a transcriptional activator to regulate the formation of antibiotics.
3. A afsR-p gene encoding AfsR-p according to claims 1 or 2.
4. The afsR-p gene according to claim 3, which has a DNA sequence of SEQ ID NO: 1.
5. The afsR-p gene according to claim 4, which is derived from Streptomyces peucetius.
6. A recombinant vector containing the gene according to claim 3.
7. A microorganism transformed by introducing the recombinant vector according to claim 6 to a host cell selected from the group consisting of bacteria, mold, and yeast.
8. The transformed microorganism according to claim 7, wherein the host cell is the genus Streptomyces having the ability of antibiotic production.
9. Primer set for identifying afsR-p gene, which has DNA sequences of SEQ ID NOs: 5 and 6.
PCT/KR2005/001800 2004-08-23 2005-06-14 Transcriptional activator derived from streptomyces peucetius and the gene thereof WO2006022476A1 (en)

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US6197591B1 (en) * 1998-09-14 2001-03-06 Pfizer Inc. Streptomyces avermitilis regulatory genes for increased avermectin production

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Title
HORINOUCHI S. ET AL: "AfsR as an integrator of signals that are sensed by multiple serine/threonine kinases in Streptomyces coelicolor A3(2).", J IND MICROBIOL BIOTECHNOL, vol. 30, no. 8, August 2003 (2003-08-01), pages 462 - 467 *
MADDURI K. ET AL: "Functional characterization and transcriptional analysis of the dnrR1 locus,which controls daunorubicin biosynthesis in Streptomyces peucetius.", J BACTERIOL, vol. 177, no. 5, March 1995 (1995-03-01), pages 1208 - 1215 *
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