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Record identifier : 567826
Personal Name - Primary Intelectual Responsibility : Matheson, Stephanie Lynne
Title and statement of responsibility : Preparatory studies towards the identification of genes regulated by the Ner-like protein (Nlp) of Escherichia coli [Thesis]
Publication, Distribution,Etc. : McGill University (Canada),, 1999
Language of the Item : eng
Dissertation of thesis details and type of degree : M.Sc.
Body granting the degree : , McGill University (Canada),
Summary or Abstract : A lacZ transcriptional fusion library was constructed without the cloned gene of interest ( i.e. nlp ). To do this, the nlp gene was interrupted using luxAB and a tetracycline resistance gene in the ? lac strain E. coli 40. This strain (LF20300) was subsequently transformed with an expression vector possessing the ara P BAD promoter and a chloramphenicol resistance marker (pBAD18Cm). The plasmid-containing strain was then lysogenized with a Mu d1 phage containing the lacZYA reporter gene and an ampicillin resistance marker. The resulting lysogens were selected on TCMG plates containing 0.2 glucose, tetracycline, chloramphenicol, and ampicillin. These lysogens were then mastered onto TCMG plates containing either 0.2 glucose or 0.2 arabinose plus X-gal and the appropriate antibiotics. Differences in blue colour were noted for eight different clones on glucose in comparison to their growth on arabinose. Total genomic DNA was isolated from each arabinose-responsive clone, digested with several different restriction endonucleases, and subjected to Southern hybridization (using labeled lacZ as a probe). Putative restriction maps were determined for those clones with single Mu d1 inserts, and used to identify candidate genes from the complete restriction map of E. coli . (Abstract shortened by UMI.).
Topical Name Used as Subject : Microbiology, Molecular biology
Information of biblio record : TL
 
 
 
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