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Isolation And Identification Of Aniline-Degrading Strain GXA7 And Cloning Of Its Aniline Dioxygenase Gene Cluster

Posted on:2005-06-05Degree:MasterType:Thesis
Country:ChinaCandidate:H K BiFull Text:PDF
GTID:2120360122498566Subject:Microbiology
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A bacterial strain GXA7, which was able to grow on aniline as a sole carbon, nitrogen and energy sources, was isolated from the activated sludge drained from the wastewater polluted by aniline for a long time. The optimum temperature and pH for its growth and aniline degradation were 32C~36C and 7.0, respectively. In suitable conditions strain GXA7 can degrade aniline with 2500mg/L at most. Based on phenotypic identification and 16S rDNA sequence analysis, strain GXA7 was identified as Delftia sp.. With specific primers designed, PCR products atd788 and atd762 were amplified by PCR. The two fragments were sequenced and closely related to aniline dioxygenase gene of Acinetobacter sp. (Accession number: D86080) with 93% and 98% overall similarity respectively. A genomic library of strain GXA7 was constructed in the cosmid vector pLAFR3 using E.coli EPIlOO as the host strain. Two recombinants, pGXA240 and pGXA310 were identified from the genomic library using in situ hybridization and Southern blotting probed with the PCR product atd788. By the analysis of subclone sequencing, we made certain that the aniline dioxygenase gene cluster of strain GXA7 was cloned.
Keywords/Search Tags:aniline, biodegradation, Delftia sp., genomic library, aninline dioxygenase gene
PDF Full Text Request
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