• Korean Journal of Microbiology
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• v.35 no.1
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• pp.13-17
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• 1999

Shigella sonnez is important causes of human enleric infcctions. S. sonnei KNIH104S was isolated from patient of shigellosis in Korea and previously reported. We cloned 1.7 kb BamHI fragment containing the asd gene encoding an aspartate $\beta$-semialdehyde dehydrogenase from chromosomal DNA of S. sonnei KNIH104S. This recombinant plasmid was named as pSAB17. E. coli $\chi$6097, an a d mutant, cannol grow on the LB medium without DL-$\alpha$, $\varepsilon$-diaminopimclic acid (50 pgiml) but E. coli x 6097(pSAB17) can grow on the same medium. We sequenccd the asd gene ol Shigella for the first time. The asd gcne was composed of 1,104 base pairs with ATG initiation codon and TAA termination codon. Sequence comparison of the asd gene exhibited 99.9% nucleolide sequence hornology with that of E. coli. Also, We constructed the balanced-lethal vector using pBluescrip SK(+) and asd gene of S. sonnei KNIH104S.

• BMB Reports
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• v.32 no.5
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• pp.436-439
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• 1999

Shigella sonnei is an important cause of human enteric infections. S. sonnei KNIH104S was previously reported to be isolated from Korean shigellosis patients. We cloned a 2.8-kb KpnI fragment containing the recA gene encoding a recombinase from the chromosomal DNA of S. sonnei KNIH104S. This recombinant plasmid was named pRAK28. E. coli HB101, a recA mutant, cannot grow on Luria-Bertani medium in the presence of the alkylating agent methylmethane sulfonate, however, E. coli HB101 harboring pRAK28 was found to grow on this medium. As far as we know, we are the first to sequence the recA gene from S. sonnei. This gene is composed of 1062 base pairs with an ATG initiation codon and a TAA termination codon. Nucleotide sequence comparison of the S. sonnei recA gene exhibited 99.7% and 99.5% identity with those of S. flexneri and E. coli, respectively.

• Proceedings of the Microbiological Society of Korea Conference
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• 2000.05a
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• pp.58-65
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• 2000

Diarrheal diseases are a major cause of both illness and death in developing countries and are caused by rotavirus, Shigella spp., Salmonella spp., enterotoxigenic Escherichia coli (ETEC), and Vibrio spp. In this study, for the development of vaccine against diarrheal diseases caused by Shigella sonei, Salmonella typhimurium, E. coli O157, and Vibrio cholerae, cloning and nucleotide sequence analysis of genes and characteristics of their gene products in E. coli were performed. For construction of attenuated strain of S. sonnei KNIH104 and Salmonella typhimurium KNIH100, the aroA genes were cloned, respectively. The recombinant plasmid $_pJP{\Delta}A45$ containing aroA deleted region and suicide vector $(_pJP5603)$ was constructed. The aroA gene deleted mutants were constructed using this recombinant plasmid. For cloning gene encoding antigenic region of E. coli O157 KNIH317, the O-antigen synthesis gene cluster and sit gene was cloned. The E. coli XL1-Blue cells harboring this recombinant plasmid showed cytotoxicity in Vero cells. The ctx gene was cloned for tile purpose of antigenic region against V. cholerae KNIH002. Sequence analysis confirmed that the virulence gene cassette was consisted of ace, zot, ctxA and ctxB genes.