Isolation, Cloning and Expression of the Genes for Microbial Polyurethane Degradation

Two fungi species, HAFB-2F-Br and HAFB-2F-Wh, have been isolated on polyurethane paint selective medium from Hill AFB paint waste samples. The mix removes polyurethane paint from the surface of painted plastic balls in less than 68 hours. Separately, HAFB-2F-Br strips the paint in 16 to 18 hours. Cloning has therefore begun with HAFB-2F-Br. Proteins have been extracted. The DNA vector and fungal host have been received. The pSV50 has been isolated and digested with Bgl II to remove the cos site and to reduce the number of Bgl II sites to one. The extracted linear plasmid DNA was ligated to itself to form a circular plasmid. Competent cells were prepared and the modified pSV50 was transformed into the competent cells. Several procedures have been used to extract the genomic DNA: the SDS approach was successful. Work is proceeding on further purification of the HAFB-2F-Br genomic library and plasmid isolation.