Molecular cloning of the arginine biosynthetic genes from Corynebacterium glutamicum
- Authors
- Chun J.-Y.; Jung S.-I.; Ko S.-Y.; Park M.-Y.; Kim S.-Y.; Lee H.-S.; Cheon C.-I.; Min K.-H.; Lee M.-S.
- Issue Date
- Dec-1996
- Publisher
- The Microbiological Society of Korea
- Citation
- Journal of Microbiology, v.34, no.4, pp 355 - 362
- Pages
- 8
- Journal Title
- Journal of Microbiology
- Volume
- 34
- Number
- 4
- Start Page
- 355
- End Page
- 362
- URI
- https://scholarworks.sookmyung.ac.kr/handle/2020.sw.sookmyung/150602
- ISSN
- 1225-8873
1976-3794
- Abstract
- Complementation cloning of the argC, E, B, D, F, and G genes in Corynebacterium glutamicum was done by transforming the genomic DNA library into the corresponding arginine auxotrophs of Escherichia coli. Recombinant plasmids containing 6.7 kb and 4.8 kb fragments complementing the K coli argB mutant were also able to complement the E. coli argC, E, A, D, and F mutants, indicating the clustered organization of the arginine biosynthetic genes within the cloned DNA fragments. The insert DNA fragments in the recombinant plasmids, named pRB1 and pRB2, were physically mapped with several restriction enzymes. By further subcloning the entire DNA fragment containing the functions and by complementation analysis, we located the arg genes in the order of ACEBDF on the restriction map. We also determined the DNA nucleotide sequence of the fragment and report here the sequence of the argB gene. When compared to that with the mutant strain, higher enzyme activity of N-acetylglutamate kinase was detected in the extract of the mutant carrying the plasmid containing the putative argB gene, indicating that the plasmid contains a functional argB gene. Deduced amino acid sequence of the argB gene shows 45%, 38%, and 25% identity to that from Bacillus strear other mophilus, Bacillus substilus, and E. coli respectively. Our long ter
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