Previous Article
J Bacteriol, May 1998, p. 2796-2799, Vol. 180, No. 10
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Cloning and Physical Mapping of the
EcoRI Fragments of the Giant Linear Plasmid
SCP1
Matthias
Redenbach,1,2
Kazuya
Ikeda,1
Masayuki
Yamasaki,1 and
Haruyasu
Kinashi1,*
Department of Molecular Biotechnology,
Graduate School of Engineering, Hiroshima University,
Higashi-Hiroshima 739-8527, Japan,1 and
Genome Research Unit, Department of Genetics,
Kaiserslautern University, D-67653 Kaiserslautern,
Germany2
Received 25 November 1997/Accepted 10 March 1998
 |
ABSTRACT |
A cosmid library was constructed for the 350-kb giant linear
plasmid SCP1 and aligned on a successive linear map. Only a
0.8-kb gap has remained uncloned in the terminal inverted repeats close to both ends. Partial digestion of the aligned cosmids
with EcoRI and hybridization with the flanking fragments of
the vector enabled physical mapping of all of the
EcoRI fragments. On this map, the methylenomycin
biosynthetic gene cluster, the insertion sequence IS466,
and the sapCDE genes coding for spore-associated proteins were localized.
| |
TEXT |
Linear plasmids have been found in a
variety of eucaryotic species, but have rarely been identified in
procaryotic organisms. However, the filamentous soil bacteria
Streptomyces spp. possess frequently linear plasmids, which
seem therefore to be a significant feature of this genus
(12). The plasmids vary in size between 12 kb
(28) and 1 Mb (24a). They contain terminal
inverted repeats (TIRs) which range between 44 bp in SLP2 of
Streptomyces lividans (3) and 95 kb in pPZG101 of
Streptomyces rimosus (6). The 5' ends of all of
the plasmids investigated were shown to be blocked by a protein.
Lin et al. (20) first demonstrated that S. lividans has an 8-Mb linear chromosome. The linear chromosome
topology has been proved for Streptomyces griseus
(19), Streptomyces ambofaciens (18),
Streptomyces coelicolor A3(2) (26), and S. rimosus (25). The close structural similarities between
Streptomyces chromosomes and linear plasmids suggest a
hypothetical idea that linear plasmids were generated by recombination
of linear chromosomes, or circular chromosomes were linearized by
integration of a linear plasmid.
SCP1 from S. coelicolor A3(2) is genetically the
best-studied linear plasmid. The element long remained unisolated but
was postulated on the basis of classical genetic analysis
(27). It was shown to carry genes for the biosynthesis of
and resistance to the antibiotic methylenomycin (2, 17).
With the introduction of pulsed-field gel electrophoresis (PFGE), SCP1
was revealed to be a 350-kb giant linear plasmid containing 80-kb-long
TIRs (13, 15).
Considerable attention has been paid to SCP1 because of its
interaction with the host chromosome. Freely replicating SCP1 mobilizes chromosomal markers randomly, as seen for all fertility plasmids in Streptomyces. In addition, Hopwood and
colleagues isolated mutants carrying an SCP1-chromosome hybrid
structure which promotes a directional DNA transfer during conjugation
(9, 10). Free SCP1-prime plasmids containing a certain
chromosomal DNA stretch were found as well as mutants
possessing SCP1 integrated in a central region of the
S. coelicolor chromosome. Those SCP1-integrated strains showed either unidirectional or bidirectional DNA
transfer with respect to the SCP1 integration site. Preliminary
structural analysis of the integrated copies of SCP1 has been carried
out for S. coelicolor A3(2) strain 2612 (an NF strain)
(8) and strains A608 (a pabA donor) and A634 (an
NF-like donor strain) (16).
To reveal in more detail the genetic organization of SCP1 and its role
in conjugative DNA transfer, we established an ordered contig of cosmid
clones covering most parts of the element.
Generation of an ordered cosmid library.
A cosmid library was
prepared for total DNA of S. coelicolor 1147, a wild-type
A3(2) strain, which carries both SCP1 and a 31-kb circular plasmid,
SCP2 (22). Total DNA was isolated and partially
digested with Sau3AI to an average size of 40 to 60 kb.
As described previously (26), the DNA was ligated to
the vector Supercos-1 (5) (Stratagene, La
Jolla, Calif.) and packaged into lambda phages which were
subsequently used to transfect Escherichia coli
Sure. Two thousand cosmid clones representing the total genome of
S. coelicolor A3(2) were screened with the SCP1 DNA, which was isolated by contour-clamped homogeneous electric fields (CHEF) (4) and nonradioactively labeled with dig-11-dUTP
(Boehringer Mannheim, Mannheim, Germany). A total of 192 clones which
hybridized strongly with the SCP1 DNA were obtained.
Supercos-1 possesses T3 and T7 promoters flanking the
BamHI cloning site. DNAs of 25 randomly selected
clones were isolated and restricted with SalI. The
dig-11-dUTP-labeled transcripts of the cosmid ends were generated from
the SalI-digested DNAs by using T3 and T7 RNA polymerases.
The labeled end probes were hybridized against colony blots of SCP1
clones to identify neighboring cosmid clones. Four additional rounds of
hybridizations, including a total of 30 cosmids, were performed to
obtain 11 cosmids which could be unambiguously aligned and cover most
parts of SCP1 (Fig. 1). Cosmid 31 is
completely located within the TIR and was therefore used for the
alignment at both ends of SCP1. It was not possible to find a clone
among the 192 SCP1 cosmids that reaches closer than 5.0 kb to the very
end, because no cosmid hybridized with plasmid pSCP201, which contains
the 4.1-kb end SpeI fragment of SCP1 (14).
View larger version (11K):
[in this window]
[in a new window]
|
FIG. 1.
Ordered cosmid and restriction maps of SCP1. In the
upper part, 12 cosmids covering almost the entire region of SCP1 are
aligned on a continuous linear map. The open and solid circles indicate
T3 and T7 promoters, respectively. pSCP201, which carries the 4.1-kb
terminal SpeI fragment of SCP1 (14), is included
at both ends. A gap has remained uncloned between pSCP201 and cosmid
31. The restriction map of SCP1 is drawn in the center. Sites for
EcoRI and rare cutting enzymes are shown above and below the
SCP1 line. Previous sequence analysis of pSCP201 revealed that five
EcoRI sites are present in the 342 nucleotides at the end of
SCP1. Among them, four EcoRI sites, the total size of whose
fragments is 0.2 kb, were omitted here to avoid complexity, but they
are included in Fig. 3A. The sizes of fragments are indicated in
kilobases. The positions of the mmr, IS466, and
sapCDE genes and two TIRs, left (TIR-L) and right (TIR-R),
are shown at the bottom. The transcription direction of the
mmr gene (24) is indicated by an arrow. Af,
AflII; As, AseI; V, EcoRV; Sp,
SpeI; Ss, SspI; X, XbaI.
|
|
Construction of the EcoRI fragment map.
To
establish a precise restriction map of SCP1, we chose EcoRI
as an appropriate enzyme. EcoRI generated a suitable number (about 40) of observable SCP1 fragments and could excise the insert from the vector. Since SCP1 and Supercos-1 contain only two
AseI sites, the 4.3- and 0.9-kb
AseI-EcoRI fragments of the latter could be used
as probes to determine the order of EcoRI fragments. Each
cosmid clone was first completely digested with AseI and then partially digested with EcoRI. The partial digest was
separated by CHEF gel electrophoresis and hybridized with the 4.3- and
0.9-kb AseI-EcoRI probes separately. This method
also confirmed the direction of the insert, because the 4.3- and 0.9-kb
fragments carry the T7 and T3 promoters, respectively.
In Fig.
2, the analysis of cosmid 53 is
shown as an example. The complete restriction digest of cosmid 53 with
AseI and
EcoRI
gave seven fragments (Fig.
2A):
four from the SCP1 DNA (14.0,
13.0, 8.6, and 6.5 kb) and three from the
vector DNA (4.3, 1.7,
and 0.9 kb). Hybridization of the 4.3-kb
AseI-
EcoRI probe to the
partial
EcoRI
digest revealed bands of 4.3, 13, 27, 33, 46, and
47 kb (Fig.
2B). This
result determined the order of
EcoRI fragments
from the T7
promoter (right-handed direction) as (4.3)-8.6-14.0-6.5-13.0-(0.9
kb). The same analysis was carried out with the 0.9-kb
AseI-
EcoRI
probe, which showed hybridizing bands
of 0.9, 14, 20, 34, 43,
and 47 kb (Fig.
2C). The order in the opposite
direction from
the T3 promoter agreed with the result described above.
View larger version (76K):
[in this window]
[in a new window]
|
FIG. 2.
Restriction analysis of cosmid 53. (A) Conventional
agarose gel electrophoresis of the complete digest of SCP1 with
AseI and EcoRI. (B and C) Southern hybridization
of the partial digest of SCP1. The SCP1 DNA was completely digested
with AseI and then partially digested with 3 U of
EcoRI in 20 µl of reaction buffer for 10 and 20 min. The
restricted DNAs were separated by CHEF gel electrophoresis and
transferred to nylon membranes. Hybridization was carried out with two
fragments of the vector as a probe: the 4.3-kb
AseI-EcoRI fragment (B) and the 0.9-kb
AseI-EcoRI fragment (C). (D) Determination of the
real size of the rightmost EcoRI fragment of cosmid 53. The
SCP1 DNA was digested with EcoRI, separated by conventional
agarose gel electrophoresis, and probed with the 13.0-kb right-end
fragment of cosmid 53. The EcoRI digests of cosmids 53 and
31 were used as references. M1,  DNA digested with
HindIII; M2, DNA-Mono Cut Mix (New England Biolabs,
Inc., Beverly, Mass.); E, EcoRI; As, AseI.
|
|
The sizes (8.6 and 13.0 kb) of two
EcoRI fragments located
at the left and right ends of cosmid 53 did not reflect real sizes,
because both fragments had been cut by
Sau3AI before
cloning.
The leftmost fragment was detected in the
EcoRI
digest of the
left-hand cosmid 63 and was determined to be 10.5 kb long
(data
not shown). The size of the rightmost fragment was deduced to
be
14.0 kb by hybridization of the 13.0-kb right-end fragment
of cosmid 53 to the complete
EcoRI digest of SCP1 itself (Fig.
2D). This
was confirmed by direct observation of the identical
fragment in the
EcoRI digest of cosmid 11 due to the TIR (data
not shown).
All of the aligned cosmids except cosmid 32 were analyzed
in this way,
and their
EcoRI fragment maps were constructed.
Cosmid 32 contains one
AseI site but no
EcoRI
site; thus, a large
EcoRI fragment extends over three
cosmids, 17, 32, and 39.
This large fragment was directly observed by
CHEF gel electrophoresis
of the
EcoRI digest of SCP1 and was
determined to be 66 kb long
(data not shown).
Although cosmid 31 carries a DNA fragment of the TIR region, it does
not reach the 4.1-kb end
SpeI fragment cloned in plasmid
pSCP201 (
14). A detailed restriction map at the end of SCP1
was constructed by using pSCP201 and cosmid 31, which revealed
that the
size of the gap between these clones was 0.8 kb (Fig.
3A). To close the gap, we tried to clone
the following five fragments
by using three cloning vectors, pUC19,
pBR322, and pACYC184, with
different copy numbers: the 1.6-kb
SpeI-
EcoRI fragment, the 2.3-kb
EcoRI
fragment, the 3.8-kb
SpeI-
EcoRV fragment, the
7.6-kb
XbaI-
EcoRV
fragment, and the 20-kb
BamHI fragment (Fig.
3A). However, we
have not succeeded in
cloning any of these fragments. This suggests
the possibility that the
region around the 0.8-kb gap has a special
structure unclonable in any
of the three vectors or that its protein
product has a deteriorative
effect on the host.
View larger version (15K):
[in this window]
[in a new window]
|
FIG. 3.
Detailed restriction maps of three regions of SCP1. (A)
Terminal region of SCP1. Four EcoRI sites at the extreme end
of SCP1 are included in this map. The gap between the SpeI
site at the right end of pSCP201 and the Sau3AI site at the
left end of cosmid 31 was deduced from this map to be 0.8 kb. (B)
mmy region. The positions of the mmr gene and
plasmid pIJ518 are shown. The map direction is opposite to that of
Chater and Bruton (2). (C) Location of IS466, the
right TIR (TIR-R), and the sapCDE genes. E,
EcoRI; Ba, BamHI; Cl, ClaI; Nd,
NdeI; Sp, SpeI; Ss, SspI; V,
EcoRV; X, XbaI.
|
|
As a result, the entire region of the SCP1 DNA except for two 0.8-kb
gaps close to both ends has been cloned, and all of the
EcoRI fragments were aligned on a linear physical map
as shown
in Fig.
1. We previously reported a map of
EcoRV fragments of
SCP1 (
13). (The number of the
EcoRV fragments was corrected
to 16, because the 20-kb band
was revealed to be a doublet.) All
of the
EcoRV sites were
now located precisely on a linear map
by restriction analysis of the
ordered cosmids. In addition, all
of the recognition sites for
AflII,
AseI,
SpeI,
SspI,
and
XbaI
were localized on the same map. The total size of
the
EcoRI fragments
was calculated to be 363.1 kb, which was
a little bit larger than
the reported size (350 kb) of the intact SCP1
(
13). We again
tested the size of the intact SCP1 by
using
Saccharomyces cerevisiae AB972 chromosomes as
size markers. The intact SCP1 moved at the
same rate with chromosome
III (data not shown), whose size is
350 kb according to Link and Olson
(
21).
Location of the mmy, IS466, and
sapCDE genes.
Several genes were reported to be
present on SCP1. Chater and Bruton (2) constructed the
restriction maps for the methylenomycin biosynthetic gene
(mmy) clusters on plasmids SCP1 and pSV1; the latter was
suggested to be a 170-kb circular plasmid (1), in contrast
to the linear structure of the former. Hybridization experiments with
pIJ518 as a probe, which carries the 7.5-kb mmy fragment of
SCP1 (2) (Fig. 3B), revealed that the resistance gene
(mmr) is located on the 10-kb EcoRI fragment of
cosmid 73. The restriction map of the mmy region on cosmid
73 agreed with that of Chater and Bruton (2).
We previously reported that the insertion sequence IS
466
(
11) is present just at the inside end of the right TIR of
SCP1
(
14). Analysis of cosmid 63 determined its location on
the
EcoRI
map as shown in Fig.
3C. Guijarro et al.
(
7) isolated spore-associated
proteins SapA and SapB from
S. coelicolor A3(2). The
sapA gene
coding for the
former protein has been cloned (
7) and was located
on
AseI fragment F of the chromosome (
26). McCormick
et al.
(
23) cloned additional
sap genes,
sapCDE, which, however, seemed
not to be essential for spore
formation, because they were mapped
to SCP1, and SCP1
strains show no effect on sporulation. Two sets of the
sapCDE genes were located in each of both TIR regions, and
their restriction
maps were constructed (
22a). Analysis of
cosmid 53 could locate
the
sapCDE genes on the
EcoRI map (Fig.
3C).
Ordered clone libraries have proved to be an excellent tool for
analysis of large genomic DNAs. For the 8-Mb
S. coelicolor A3(2) chromosome, an ordered cosmid library has been established
and
used extensively for gene mapping (
27), and it is now being
used for the genome project in collaboration with the Sanger Centre
and
the John Innes Centre. Here we reported the generation of
an ordered
cosmid contig of the giant linear plasmid SCP1, leaving
only two 0.8-kb
gaps uncloned. A detailed
EcoRI restriction map
was
constructed, and the genetic markers IS
466,
mmy, and
sapCDE were assigned to specific SCP1
fragments. These results will be
of great value for study of the
structure and function of SCP1
and analysis of the interaction of
SCP1 with the chromosome of
S. coelicolor A3(2).
| |
ACKNOWLEDGMENTS |
We thank Keith Chater for plasmid pIJ518; John Cullum for
pMT644, which carries IS466; and Joe McCormick for
pRC2 and pRD2, which carry a part of sapC and
sapD, respectively.
M. Redenbach has been supported by a scholarship from the Japan Society
for the Promotion of Sciences.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Department of
Molecular Biotechnology, Graduate School of Engineering, Hiroshima
University, Higashi-Hiroshima 739-8527, Japan. Phone: 81-824-7869. Fax:
81-824-24-7869. E-mail:
kinashi{at}ipc.hiroshima-u.ac.jp.
| |
REFERENCES |
| 1.
|
Aguilar, A., and D. A. Hopwood.
1982.
Determination of methylenomycin A synthesis by the pSV1 plasmid from Streptomyces violaceus-ruber SANK95570.
J. Gen. Microbiol.
128:1893-1901[Abstract/Free Full Text].
|
| 2.
|
Chater, K. F., and C. J. Bruton.
1985.
Resistance, regulatory and production genes for the antibiotic methylenomycin are clustered.
EMBO J.
4:1893-1897[Medline].
|
| 3.
|
Chen, C. W.,
T.-W. Yu,
Y.-S. Lin,
H. M. Kieser, and D. A. Hopwood.
1993.
The conjugative plasmid SLP2 of Streptomyces lividans is a 50 kb linear molecule.
Mol. Microbiol.
7:925-932[Medline].
|
| 4.
|
Chu, G.,
D. Vollrath, and R. W. Davis.
1986.
Separation of large DNA molecules by contour-clamped homogeneous electric fields.
Science
234:1582-1585[Abstract/Free Full Text].
|
| 5.
|
Evans, G. A.,
K. Lewis, and B. E. Rothenberg.
1989.
High efficiency vectors for cosmid microcloning and genomic analysis.
Gene
79:9-20[Medline].
|
| 6.
|
Gravius, B.,
D. Glocker,
J. Pigac,
K. Pandza,
D. Hranueli, and J. Cullum.
1994.
The 387 kb linear plasmid pPZG101 of Streptomyces rimosus and its interactions with the chromosome.
Microbiology
140:2271-2277[Abstract/Free Full Text].
|
| 7.
|
Guijarro, J.,
R. Santamaria,
A. Schauer, and R. Losick.
1988.
Promoter determining the timing and spatial localization of transcription of a cloned Streptomyces coelicolor gene encoding a spore-associated polypeptide.
J. Bacteriol.
170:1895-1901[Abstract/Free Full Text].
|
| 8.
|
Hanafusa, T., and H. Kinashi.
1992.
The structure of an integrated copy of the giant linear plasmid SCP1 in the chromosome of Streptomyces coelicolor 2612.
Mol. Gen. Genet.
231:363-368[Medline].
|
| 9.
|
Hopwood, D. A., and T. Kieser.
1993.
Conjugative plasmids of Streptomyces, p. 293-311.
In
D. B. Clewell (ed.), Bacterial conjugation. Plenum Press, New York, N.Y.
|
| 10.
|
Hopwood, D. A., and H. M. Wright.
1976.
Interaction of the plasmid SCP1 with the chromosome of Streptomyces coelicolor A3(2), p. 607-619.
In
K. D. MacDonald (ed.), Second International Symposium on the Genetics of Industrial Microorganisms. Academic Press, London, United Kingdom.
|
| 11.
|
Kendall, K., and J. Cullum.
1986.
Identification of a DNA sequence associated with plasmid integration in Streptomyces coelicolor A3(2).
Mol. Gen. Genet.
202:240-245[Medline].
|
| 12.
|
Kinashi, H.
1994.
Linear plasmids from actinomycetes.
Actinomycetologica
8:87-96.
|
| 13.
|
Kinashi, H., and M. Shimaji-Murayama.
1991.
Physical characterization of SCP1, a giant linear plasmid from Streptomyces coelicolor.
J. Bacteriol.
173:1523-1529[Abstract/Free Full Text].
|
| 14.
|
Kinashi, H.,
M. Shimaji-Murayama, and T. Hanafusa.
1991.
Nucleotide sequence analysis of the unusually long terminal inverted repeats of giant linear plasmid, SCP1.
Plasmid
26:123-130[Medline].
|
| 15.
|
Kinashi, H.,
M. Shimaji, and A. Sakai.
1987.
Giant linear plasmids in Streptomyces which code for antibiotic biosynthesis genes.
Nature
328:454-456[Medline].
|
| 16.
|
Kinashi, H.,
M. Murayama,
H. Matsushita, and O. Nimi.
1993.
Structural analysis of the giant linear plasmid SCP1 in various Streptomyces coelicolor strains.
J. Gen. Microbiol.
139:1261-1269.
|
| 17.
|
Kirby, R., and D. A. Hopwood.
1977.
Genetic determination of methylenomycin synthesis by the SCP1 plasmid of Streptomyces coelicolor A3(2).
J. Gen. Microbiol.
98:239-252[Abstract/Free Full Text].
|
| 18.
|
Leblond, P.,
G. Fischer,
F.-X. Francou,
F. Berger,
M. Guerineau, and B. Decaris.
1996.
The unstable region of Streptomyces ambofaciens includes 210-kb terminal inverted repeats flanking the extremities of the linear chromosomal DNA.
Mol. Microbiol.
19:261-271[Medline].
|
| 19.
|
Lezhava, A.,
T. Mizukami,
T. Kajitani,
D. Kameoka,
M. Redenbach,
H. Shinkawa,
O. Nimi, and H. Kinashi.
1995.
Physical map of the linear chromosome of Streptomyces griseus.
J. Bacteriol.
177:6492-6498[Abstract/Free Full Text].
|
| 20.
|
Lin, Y.-S.,
H. M. Kieser,
D. A. Hopwood, and C. W. Chen.
1993.
The chromosomal DNA of Streptomyces lividans 66 is linear.
Mol. Microbiol.
10:923-933[Medline].
|
| 21.
|
Link, A. J., and M. V. Olson.
1991.
Physical map of the Saccharomyces cerevisiae genome at 110-kilobase resolution.
Genetics
127:681-698[Abstract].
|
| 22.
|
Lydiate, D. J.,
F. Malpartida, and D. A. Hopwood.
1985.
The Streptomyces plasmid SCP2*: its functional analysis and development into useful cloning vectors.
Gene
35:223-235[Medline].
|
| 22a.
| McCormick, J. R., and R. Losick. Personal
communication.
|
| 23.
|
McCormick, J. R.,
R. Santamaria, and R. Losick.
1991.
In
The genes for three spore-associated proteins are encoded on linear plasmid SCP1 in Streptomyces coelicolor, abstr. P1-125. Abstracts of the 8th International Symposium on Biology of Actinomycetes, Madison, Wis
.
|
| 24.
|
Neal, R. J., and K. F. Chater.
1987.
Nucleotide sequence analysis reveals similarities between proteins determining methylenomycin A resistance in Streptomyces and tetracycline resistance in eubacteria.
Gene
58:229-241[Medline].
|
| 24a.
| Pandza, K., and J. Cullum. Personal communication.
|
| 25.
|
Pandza, K.,
G. Pfalzer,
J. Cullum, and D. Hranueli.
1997.
Physical mapping shows that the unstable oxytetracycline gene cluster of Streptomyces rimosus lies close to one end of the linear chromosome.
Microbiology
143:1493-1501[Abstract/Free Full Text].
|
| 26.
|
Redenbach, M.,
H. M. Kieser,
D. Denapaite,
A. Eichner,
J. Cullum,
H. Kinashi, and D. A. Hopwood.
1996.
A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2).
Mol. Microbiol.
21:77-96[Medline].
|
| 27.
|
Vivian, A.
1971.
Genetic control of fertility in Streptomyces coelicolor A3(2): plasmid involvement in the interconversion of UF and IF strains.
J. Gen. Microbiol.
69:353-364.
|
| 28.
|
Wu, X., and K. L. Roy.
1993.
Complete nucleotide sequence of a linear plasmid from Streptomyces clavuligerus and characterization of its RNA transcripts.
J. Bacteriol.
175:37-52[Abstract/Free Full Text].
|
J Bacteriol, May 1998, p. 2796-2799, Vol. 180, No. 10
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Arakawa, K., Mochizuki, S., Yamada, K., Noma, T., Kinashi, H.
(2007). {gamma}-Butyrolactone autoregulator-receptor system involved in lankacidin and lankamycin production and morphological differentiation in Streptomyces rochei. Microbiology
153: 1817-1827
[Abstract]
[Full Text]
-
Ramos, J. L., Martinez-Bueno, M., Molina-Henares, A. J., Teran, W., Watanabe, K., Zhang, X., Gallegos, M. T., Brennan, R., Tobes, R.
(2005). The TetR Family of Transcriptional Repressors. Microbiol. Mol. Biol. Rev.
69: 326-356
[Abstract]
[Full Text]
-
Yamasaki, M., Kinashi, H.
(2004). Two Chimeric Chromosomes of Streptomyces coelicolor A3(2) Generated by Single Crossover of the Wild-Type Chromosome and Linear Plasmid SCP1. J. Bacteriol.
186: 6553-6559
[Abstract]
[Full Text]
-
Shi, L., Zhang, W.
(2004). Comparative analysis of eukaryotic-type protein phosphatases in two streptomycete genomes. Microbiology
150: 2247-2256
[Abstract]
[Full Text]
-
Stecker, C., Johann, A., Herzberg, C., Averhoff, B., Gottschalk, G.
(2003). Complete Nucleotide Sequence and Genetic Organization of the 210-Kilobase Linear Plasmid of Rhodococcus erythropolis BD2. J. Bacteriol.
185: 5269-5274
[Abstract]
[Full Text]
-
Yamasaki, M., Ikuto, Y., Ohira, A., Chater, K., Kinashi, H.
(2003). Limited regions of homology between linear and circular plasmids encoding methylenomycin biosynthesis in two independently isolated streptomycetes. Microbiology
149: 1351-1356
[Abstract]
[Full Text]
-
Gust, B., Spatz, K., Spychaj, A., Redenbach, M.
(2001). Region-Specific Transcriptional Activity in the Genome of Streptomyces coelicolor A3(2). Appl. Environ. Microbiol.
67: 3598-3602
[Abstract]
[Full Text]
-
Wang, Z.-X., Li, S.-M., Heide, L.
(2000). Identification of the Coumermycin A1 Biosynthetic Gene Cluster of Streptomyces rishiriensis DSM 40489. Antimicrob. Agents Chemother.
44: 3040-3048
[Abstract]
[Full Text]
-
Yamasaki, M., Miyashita, K., Cullum, J., Kinashi, H.
(2000). A Complex Insertion Sequence Cluster at a Point of Interaction between the Linear Plasmid SCP1 and the Linear Chromosome of Streptomyces coelicolor A3(2). J. Bacteriol.
182: 3104-3110
[Abstract]
[Full Text]
-
Steffensky, M., Mühlenweg, A., Wang, Z.-X., Li, S.-M., Heide, L.
(2000). Identification of the Novobiocin Biosynthetic Gene Cluster of Streptomyces spheroides NCIB 11891. Antimicrob. Agents Chemother.
44: 1214-1222
[Abstract]
[Full Text]
Top
Abstract
Text
