The cloning and characterization of a Butyrivibrio fibrisolvens H17c glnA regulatory element in E. coli
Butyrivibrio fibrisolvens HI 7C is an important anaerobic bacterium which occurs in the rumen of most ruminants. A key factor affecting the growth of this bacterium is the availability of nitrogen sources, particularly in the form of ammonia. The aims of this study were to attempt the isolation of a gene/genes involved in the regulation of the B.fibrisolvens type III glutamine synthetase (GS), which is a key enzyme involved in ammonia assimilation in rumen bacteria. An existing B. fibrisolvens gene bank, as well as the B. fibrisolvens glnA gene cloned onto a low 11 copy number plasmid, were used to generate a heterologous, Escherichia coli- based, two plasmid, in trans system. This system was used to isolate an E. coli clone which showed increased GS activity levels (2.5-fold) and a retarded growth rate phenotype in complete media.
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Department of Molecular and Cell Biology