Molecular Cloning of nifHD from Rhizobium sp. SNU003

Rhizobium sp. SNU003의 nifHD 클로닝

  • 강명수 (서울대학교 자연과학대학 생물학과) ;
  • 안정선 (서울대학교 자연과학대학 생물학과)
  • Published : 1993.04.01

Abstract

Genes for dinitrogenase reductase (nifH) and dinitogenase a subunit (nifD) were found to be located on 7.9 kb of EcoRI, 6.5 kb of Sail, 7.3 kb of HindlII and 4.4 kb of Pstl fragments of the genomic blot of Rhizobium sp. SNU003. a symbiotic strain from root nodule of Canavalia lineata. Nine recombinant phage nif-clones were selected from the genomic library constructed by using EMBL-3 BamHI arms of bacteriophage lambda. Among them. Rnif-6 had insert DNA of 15.3 kb. in which 7.6 kb of BamHI!SacI fragment contained nifHD region. Therefore, the 7.6 kb fragment was subcloned into pUC19 and partial restriction map was constructed. As the results, nifH and nifD were found to be located continuously on 4.5 kb of BamHI/BglIl in the genome of Rhizobium sp. SNU003 strain.

Keywords

nifH;nifD Rhizobium sp;SNU003;Canavalia lineata;genomic library