Korean Journal of Plant Tissue Culture (식물조직배양학회지)

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Expression of Helicobacter pylori Urease Subunit A in Plant

Helicobacter pylori urease subunit A 단백질의 식물체내에서의 발현

  • 이효정 (대구효성가톨릭대학교 생명자원학부) ;
  • 이만형 (대구효성가톨릭대학교 약학대학) ;
  • 신동일 (대구효성가톨릭대학교 생명자원학) ;
  • 정일경 (대구효성가톨릭대학교 생명자원학부) ;
  • 최성진 (대구효성가톨릭대학교 생명자원학부) ;
  • 박희성 (대구효성가톨릭대학교 생명자원학부)
  • Published : 2000.05.01
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Abstract

Helicobacter pylori, an etiologic agent of gastritis and peptic ulceration, produces urease which elicits a powerful immunoglobulin response in H. pylori-infected individuals. To establish a model plant vaccine agains H. pylori, 750 bp -ureA DNA amplified by polymerase chain reaction from pH 808 plasmid harboring urease gene cluster was cloned and manipulated to be expressed in tobacco plants. From the regenerated transgenic tobacco plants, ureA DNA integration,m its mRNA expression and protein synthesis were analyzed and confirmed by standard molecular techniques. The CaMV 35S promoter-driving ureA construct was expressed to produce a 30 kDa protein which was identical with bacterial UreA in size when detected on immunoblot of SDS polyacrylamide gel electrophoresis.

위염과 위궤양을 일으키는 Helicobacter pylori는 그 감염 환자에서 강한 항체형성을 유도하는 urease를 생산한다. 식물체를 이용하여 H. pylori에 대한 식용백신 모델을 제조하기 위하여 H. pylori의 urease 유전자를 지니고 있는 pH808 plasmid로부터 750bp의 ureA DNA를 PCR에 의해서 증폭한 후 이를 담배식물에서 발현이 되도록 조작하였다. 재분화된 형질전환 식물체로부터 ureA 유전자의 도입과 mRNA발현 및 단백질합성을 분석하였다. CaMV 35S promoter에 의한 ureA 유전자의 발현은 SDS polyacrylamide 전기영동 및 immunoblot실험에서 박테리아로부터의 재조합 단백질과 같은 크기의 30 kDA UreA 단백질이 생산되었음을 확인할 수 있었다.

Keywords

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