Cloning and expression of the Thiobacillus ferrooxidans glutamine synthetase gene in Escherichia coli.
The glutamine synthetase (GS) gene glnA of Thiobacillus ferrooxidans was cloned on recombinant plasmid pMEB100 which enabled Escherichia coli glnA deletion mutants to utilize (NH4)2SO4 as the sole source of nitrogen. High levels of GS-specific activity were obtained in the E. coli glnA deletion mutants containing the T. ferrooxidans GS gene. The cloned T. ferrooxidans DNA fragment containing the glnA gene activated histidase activity in an E. coli glnA glnL glnG deletion mutant containing the Klebsiella aerogenes hut operon. Plasmid pMEB100 also enabled the E. coli glnA glnL glnG deletion mutant to utilize arginine or low levels of glutamine as the sole source of nitrogen. There was no detectable DNA homology between the T. ferrooxidans glnA gene and the E. coli glnA gene.
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D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004269:DNA, Bacterial / Q000032:analysis; D015246:Deoxyribonuclease EcoRI; D015247:Deoxyribonuclease HindIII; D015252:Deoxyribonucleases, Type II Site-Specific; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005786:Gene Expression Regulation; D005974:Glutamate-Ammonia Ligase / Q000235:genetics*; D009154:Mutation; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D013855:Thiobacillus / Q000201:enzymology / Q000235:genetics*
D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes; D015246:Deoxyribonuclease EcoRI; D015247:Deoxyribonuclease HindIII; C024388:CTGCAG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; D005974:Glutamate-Ammonia Ligase