Cloning The Phenol Hydroxylase Gen From Acinetobacter Junii GXP04

A bacterial strain GXP04, which was able to grow on phenol as a sole carbon, nitrogen and energy sources, was isolated from the soil sample which was polluted by phenol for a long time. On the basis of the analysis of the 16SrDNA sequence, GXP04 was identified as Acinetobacter junii . With specific primers designed , 684bp nucleotide fragment was amplified by PCR. This fragment was sequenced and closely related to phenol hydroxylase gene of Acinetobacter radioresistens which was reported before with 97% similarity. The total DNA of GXP04 was digested with different restriction enzymes. We used labeled this 684bp nucleotide fragment as a specific probe, the phenol hydroxylase's closely related gene was got according Southern blotting. A genomic library of strain GXP04 was constructed in the cosmid vector pLAFR3. 12 recombinants, were identified from the genomic library using in situ hybridization and same probes as Southern blotting. According such as TAIL-PCR, Southern blotting ect methods, 8 recombinants were identified which including whole phenol hydroxylase gene . This phenol hydroxylase's nucleotide sequence and amino acid sequence were analysised .We analysised the basic character of this protein and laid a foundation for reseaching Phenolhydroxylase gene.