Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Cloning and Nucleotide Sequence Analysis of the aroA Gene from Salmonella typhi KNIH100

Salmonella typhi KNIH100으로부터 aroA 유전자의 클로닝과 염기서열 분석

  • 길영식 (충북대학교 자연과학대학 생명과학부) ;
  • 신희정 (충북대학교 자연과학대학 생명과학부) ;
  • 김영창 (충북대학교 유전공학연구소)
  • Published : 2000.03.01
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Abstract

Salmonella typhi is one of important causes of human enteric infections. S. typhi KNIH100 was isolated from a patient of typhoid fever in Korea. We cloned a 5.0 kb SalⅠ fragment containing the aroA gene encoding a 5-enolpyruvylshikimate-3-phosphate synthetase from chromosomal DNA of this strain. This recombinant plasmid was named pSAL80. E. coli CGSC2829, an aroA- mutant, was not grown on the M9 minimal medium but E. coli CGSC2829 (pSAL80) was grown on the M9 minimal medium. The aroA gene was composed of 1,284 base pairs with ATG initiation codon and TAA termination codon. Sequence comparison of the aroA gene exhibited 99%, 98%, and 77% identity with those of S. typhi Ty2, S. typhimurium, and E. coli respectively. As in the cases of Shigella sonnei and E. coli, the serC and aroA genes lie in a single operonic structure.

장티푸스는 Salmonella typhi에 의해 유발되는 장감염성 질환으로 사람과 동물에 공통되는 질병이다. 본 연구에서는 국립보건원과 공동연구를 수행하여 한국형 장티푸스 유발균인 S. typhi KNIH100을 분리하였다. 분리된 S. typhi KNIH100의 염색체 DNA로부터 방향족 아미노산의 생합성에 관여하는 효소인 5-enolpyruvylshikimate-3-phosphate synthetase를 암호화하는 aroA 유전자를 포함하는 약 5.0 kb의 SalI절편을 pBluescriptII SK(+) vector와 aroA 돌연변이주인 E. coli CGSC2829를 이용하여 클로닝하였다. 그리고 이 클론을 pSAL80이라 명명하였다. 클로닝된 재조합 plasmid인 pSAL80에는 ATG 개시코돈과 TGA 종결코돈을 포함하는 1,284 염기로 구성된 aroA 유전자가 위치하고 있었으며, 다른 장내세균과 마찬가지로 serC와 하나의 오페론을 구성하고 있음을 밝혔다. 또한 S. typhi Ty2, S. typhimurium, 그리고 E. coli 등 다른 장내세균의 aroA 유전자와 상동성을 비교하여 본 결과 각각 99%, 95%, 77%의 상동성을 나타내었다.

Keywords

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