دراسة وراثية عن الطوافر البكتيرية المفككة للمركبات الفينولية
Author name:
دانية منعم حامد الجادر
Supervisor name:
صبحي دواد حمزة | ميساء جاسب
General topic:
Biology
Specific topic:
Biotechnologies
Degree:
Master
University:
University of Baghdad - College Of Science - Department Of Biotechnology
Language:
Arabic
University location:
Baghdad
First pages:
24T3106 - p.pdf
Abstract:
During this study (30) bacterial isolates from (55) soil samples and (10) water samples previously contaminated with petroleum oils were isolated and then screened for their capability for degrading phenol and analine . only (5) isolates were selected which show higher growth and degradation activity on the two substrates and identified Pseudomonas aeruginosa A13 and A5 , Escherichia coli A2 and A11 ، Enterobacter cloacae A8. Adaptation of isolates (A13, A11, A8, A5, A2) in higher concentrations of phenol (500 - 3000) µg/ml and analine (5003500) µg/ml were done . As much as (70,75and 80)% of (2000 µg/ml) phenol and (70,75 and 85)% of (2500 µg/ml) analine were degraded after 3 days of incubation. Number of mutants were generated by using 500 µg/ml of Nitrosoguanidine at incubations time (0.5,1.0and 2.0) hr .It was found that period of 1hr enough to induce mutation in the bacterial isolates in this study . and increased degradation activity as much as (90,90and 97)% of (2000 µg/ml) phenol and (90,97 and 100)% of (2500 µg/ml) analine were degraded by isolates (A13M2, A11M4and A8M12)respectively . Genetic analysis by electrophoretic agarose gel show that (A13, A8 and A8M12) isolates contain three plasmid bands However isolates (A11, A11M4 and A13M2) contain only two . Curing of plasmid frome the three isolates,« A11M4, A13M2 and A8M12»,were done by using acridine orange , ethidum bromide and SDS . It was found that 2200 µg/ml SDS and200 µg/ml acridine orange were capable of curing plasmids frome A11M4, A13M2 . All attempts for curing plasmid of isolates A8M12 was failed , however transformation of plasmid DNA of isolates A11M4, A13M2 and A8M12 in to E. coli MM294 was achieved . Results of the experiments indicate that these plasmids were responsible for degradation ability in the transformed isolates.
