Title Molecular cloning of the arginine biosynthetic genes from corynebacterium glutamicum
Author Chun, Jae Yeon · Jung, Sam Il · Ko, Soon Young · Park, Mee Young · Kim, Soo Young · Lee, Heung Shickc² · Cheon, Choong Ill · Min, Kyung Hee · Lee, Myeong Sok *
Address Department of biology, Sookmyung Woomen's University; ¹Department of Biology, Kunkuk University; ²Department of Biotechnology, Korea University
Bibliography Journal of Microbiology, 34(4),355-362, 1996,
Key Words Corynebacterium glutamicum, arginine biosynthesis, N-acetylglutamate, complementation, molecular cloning
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 fo Escherichia coli. Recombinant plasmids containing 6.7 kb and 4.8kb fragments complementing the E. coli arg B 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 nutant strain, higherh enzyme activity of N-acetylglutamate kinase was detacted 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 shows45%, 38%, and 25% identity to that from Bacillus strearothermophilus, Bacillus substilus, and E. coli respectively. Our long term goal is genetically engineering C. glutamicum which produces more arginine than a wild type strain does.
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