Construction of rhizobium-E. coli shuttle vector using replication and mobilization function of indigenous multicopy plasmid from rhizobium

Rhizobium muliticopy plasmid의 복제 및 이주 기능을 이용한 rhizobium-E. coli shuttle vector 구축

  • 조무제 (경상대학교 자연과학대학 생화학과) ;
  • 신평균 (경상대학교 농과대학 농화학과) ;
  • 최영주 (경상대학교 농과대학 농화학과) ;
  • 강규영 (경상대학교 농과대학 농화학과) ;
  • 윤한대 (경상대학교 농과대학 농화학과)
  • Published : 1989.06.01

Abstract

the vector, pGUR19, for Rhizobium gene manipulation, was constructed by combining the replication and mobilization function of indigenous multicopy plasmid from Acacia(Robinia pseudoacacia L.) Rhizobia sp86 with E. coli cloning vehicle, pBR322. The vector could be efficiently mobilized by RP4 tra function incorporated into chromosome of E. coli named SM10 and efficiently transferred to various gram negative hosts including Rhizobium and Afrobacterium by transformation. Mobilization frequency of the constructed vector was ranged from $1.2\times 10^{-2}$ (E.coli HB 101) to $4.6\times 10^{-4}$ (A. tumefaciens 15955) and transformation frequency was ranged from $5.4\times 10^{-7}$(E. coli HB101) to $1.2\times 10^{-10}$ (A. tumefaciens 15955). The vector, pGUR19, was stably replicated and maintained in a variety of Rhizobium and Agrobacterium.

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