The Library
Physiological and genetic studies of deoxynojirimycin production in streptomyces species
Tools
Robson, William Nigel (1993) Physiological and genetic studies of deoxynojirimycin production in streptomyces species. PhD thesis, University of Warwick.
|
PDF
WRAP_THESIS_Robson_1993.pdf - Requires a PDF viewer. Download (20Mb) |
Official URL: http://webcat.warwick.ac.uk/record=b1416232~S15
Abstract
Genomic libraries of S. lavendulae and S. subrutilus
were constructed in S. lividans using the technique of
shotgun cloning. S. lividans is a genetically well
characterised recipient for heterologous DNA, but plasmid
deletions or the entry of plasmids with small inserts
occurred during transformation of the DNA into
S. lividans This was not due to a straightforward
restriction-modif ication system as this possibility was
checkea using the KC301 phage. Several gene libraries
were produced, using high and low copy number plasmids,
and the resultant transformants screened.
The nature of deoxynojirimycin (DNJ) and its lack of
microbial activity prevented use of a bioassay. However,
the inhibition of cc-glucosidase by DNJ was exploited by
development of a quantitative assay system f or DNJ and
nojirimycin (NOJ). The assay could not only detect DNJ
and NOJ-producing clones, but could also assess the titre
of DNJ and NOJ in culture broths. The assay was used to
demonstrate differential production of DNJ and NOJ by
selected StreRtOIFYces strains of cluster 61 (the
S-lavendulae species group). The assay also confirmed
the effectiveness of using microtitre plates as an
effective screening procedure. The microtitre screening
programme generated further data and statistical
treatment of the results delimited the number of isolates
for further examination. No DNJ-producing colony was
detected and examination of the size of the DNA inserts
showed almost all were too small to contain the DNJ gene
cluster.
Additionally, blocked DNJ production mutants were
characterised by the feeding of NOJ, one of the mutants
successfully converted NOJ to DNJ.
Item Type: | Thesis (PhD) | ||||
---|---|---|---|---|---|
Subjects: | Q Science > QR Microbiology | ||||
Library of Congress Subject Headings (LCSH): | Antibiotics -- Synthesis, Streptomyces -- Genetics, Streptomyces -- Physiology | ||||
Official Date: | September 1993 | ||||
Dates: |
|
||||
Institution: | University of Warwick | ||||
Theses Department: | Department of Biological Sciences | ||||
Thesis Type: | PhD | ||||
Publication Status: | Unpublished | ||||
Supervisor(s)/Advisor: | Wellington, E. M. H. (Elizabeth M. H.), 1954- | ||||
Sponsors: | Xenova (Firm) | ||||
Extent: | xix, 206 leaves | ||||
Language: | eng |
Request changes or add full text files to a record
Repository staff actions (login required)
View Item |
Downloads
Downloads per month over past year