Molecular cloning of lipopolysaccharide genes of the Vibrio cholerae in E. coli HB101]
霍乱弧菌脂多糖基因在大肠杆菌中的克隆

A genomic library of the V. cholerae 178 (Eltor biotype, Ogawa serotype) was constructed by using cosmid pHC 79 as a cloning vector. We screened the library with immune agglutination test and colonies solid phase ELISA. 13 positive recombinants which could express the O antigen of the V. cholerae lipopolysaccharide (LPS) were acquired. The LPS was then extracted from a positive recombinant PMM-VO 38 by using hot phenol-water method. It was found that purified LPS specifically reacted to antisomatic serum against the V. cholerae. The restriction endonucleases analysis showed that the molecular weight of the recombination cosmid PMM-VO 38 was about 46 kb.