Isolation of a gene from Burkholderia cepacia IS-16 encoding a protein that facilitates phosphatase activity

A genomic library from Burkholderia cepacia IS-16 was constructed in Escher ichia coli by partial Sau3AI digestion of the chromosomal DNA, with the pla smid vector Bluescript SK. This library was screened for clones able to gro w as green stained colonies on selective medium developed for detecting pho sphatase-positive colonies. Three green-stained clones (pFS1, pFS2, and pFS 3) carried recombinant plasmids harboring DNA inserts of 5.0, 8.0, and 0.9 kb, respectively, DNA hybridization experiments demonstrated the presence o f overlapping DNA fragments in the three clones and that these three clones were all derived from Burkholderia in cepacia IS-16 genomic DNA. DNA seque nce analysis, together with polyacrylamide gels of proteins encoded by E. c oli containing pFS3, suggested that the isolated 0.9-kb DNA fragment encode s the functional portion of a phosphate transport protein.