Expression of genes from the marine bacterium Alteromonas haloplanktis 214 in Escherichia coli K-12.

DNA from the marine bacterium Alteromonas haloplanktis 214 was partially digested with Sau 3A and inserted into the Bam HI site of the cloning vector pBR322. The ligation mixture was used to transform Escherichia coli HB101. The gene bank plasmid preparation obtained was used to transform Escherichia coli K-12 strain EO2717, an organism auxotrophic for histidine, arginine, adenine, uracil and thiamin. Prototrophic transformants for each of the five metabolites were isolated using appropriate minimal media for selection. Plasmids isolated from each of the transformants were shown by hybridization to contain A. haloplanktis DNA and to be capable of complementing the appropriate mutation in E. coli EO2717. Restriction maps showed that each of the plasmids was different.