Antibiotic production and inactivation by a producing organism
Antibiotic production and inactivation by a producing organism
Production of the enzyme aminoglycoside 3'-phosphotransferase by B. circulans has been studied and shown to occur in a development manner. Studies on the DNA encoding the phosphotransferase (APH), using the techniques of S1 mapping and reverse transcriptase primer extension, have shown that the gene has different transcription start points in E. coli and B. circulans. An inspection of the gene sequence indicated that in the case of E. coli the gene is under the control of a σ55 type promotor and that maximal production of the phosphotransferase in B. circulans is under the control of a σ37 type promotor. A 7.5kb EcoRl fragment of B. circulans DNA (containing the APH) gene has been cloned into the E. coli/B. subtilis shuttle vector pHV33 and transformed into B. subtilis to produce organism JBB101. This organism has been shown to produce the enzyme aminoglycoside 3'-phosphotransferase, and investigations into the production of the protein by this organism have indicated that this organism also expresses the APH gene in a developmental manner. A cosmid bank of B. circulans DNA has been produced using the cosmid pHC79 and from this 20 clones have been isolated which express the APH gene. The DNA in these clones surrounding the APH gene has been mapped using restriction digests and Southern blotting and found to cover approximately 50kb of the B. circulans chromosome. Filter paper matings of B. circulans and S. jaecalis have been performed and mutants of B. circulans have been produced, due to the insertion of the conjugative shuttle transposon Tn916, which are unable to produce the antibiotic butirosin. Screening of these mutants with DNA from around the APH gene, by Southern blotting, showed that in each case the mutated genes were not within an area of approximately 25kb of the APH gene.(DX86697)
University of Southampton
1988
Ball, John Matthew
(1988)
Antibiotic production and inactivation by a producing organism.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
Production of the enzyme aminoglycoside 3'-phosphotransferase by B. circulans has been studied and shown to occur in a development manner. Studies on the DNA encoding the phosphotransferase (APH), using the techniques of S1 mapping and reverse transcriptase primer extension, have shown that the gene has different transcription start points in E. coli and B. circulans. An inspection of the gene sequence indicated that in the case of E. coli the gene is under the control of a σ55 type promotor and that maximal production of the phosphotransferase in B. circulans is under the control of a σ37 type promotor. A 7.5kb EcoRl fragment of B. circulans DNA (containing the APH) gene has been cloned into the E. coli/B. subtilis shuttle vector pHV33 and transformed into B. subtilis to produce organism JBB101. This organism has been shown to produce the enzyme aminoglycoside 3'-phosphotransferase, and investigations into the production of the protein by this organism have indicated that this organism also expresses the APH gene in a developmental manner. A cosmid bank of B. circulans DNA has been produced using the cosmid pHC79 and from this 20 clones have been isolated which express the APH gene. The DNA in these clones surrounding the APH gene has been mapped using restriction digests and Southern blotting and found to cover approximately 50kb of the B. circulans chromosome. Filter paper matings of B. circulans and S. jaecalis have been performed and mutants of B. circulans have been produced, due to the insertion of the conjugative shuttle transposon Tn916, which are unable to produce the antibiotic butirosin. Screening of these mutants with DNA from around the APH gene, by Southern blotting, showed that in each case the mutated genes were not within an area of approximately 25kb of the APH gene.(DX86697)
This record has no associated files available for download.
More information
Published date: 1988
Identifiers
Local EPrints ID: 461040
URI: http://eprints.soton.ac.uk/id/eprint/461040
PURE UUID: 159642aa-c646-4754-9015-032be1c19c9d
Catalogue record
Date deposited: 04 Jul 2022 18:34
Last modified: 04 Jul 2022 18:34
Export record
Contributors
Author:
John Matthew Ball
Download statistics
Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.
View more statistics