Download Study the Genetic Basis of Some Morphological Characters for

Study the Genetic Basis of Some
Morphological Characters for
Proteus Bacteria Isolasted from Medical
Samples
A Thesis Submitted
By
Saja Dhia Al-Deen Mustafa Al-Redhwany
To
The Council of College of Education,
University of Mosul In Partial Fulfillment of
Requirements for the Degree of M.Sc. In
Biotechnology
Supervised by
Assist. Prof.
Dr. Khalid D. Ahmed
1433 A.H.
2012 A.D.
ABSTRACT
The present study includes collection of 30 bacterial isolates of
genus Proteus during two months (July and August) of 2010 from
different human infections in Mosul Hospital. These bacteria were
identified
depending
on
morphological, cultural
characters
and
biochemical tests and the results revealed that these bacteria belong to the
genus Proteus and of two species P. mirabilis and P. vulgaris with
prevalence of P. vulgaris species (21 isolates) over P. mirabilis
(9 isolates). The resistance of bacterial isolates were tested to 10 different
antibiotics, 4 heavy metals and swarming phenomenon. The results
showed that there are variation in their antibiotic resistance with range
ratio (60-70)%, to heavy metals (70-80)% and all isolated display
swarming phenomenon.
Characterization of plasmid DNA content for five bacterial isolates
belonging to P. mirabilis and P. vulgaris were carried out by using
agarose gel electrophorasis and the results showed isolate the presence of
discrete clear and isolated band in each species and of similar size present
in agarose gel, which indicate that one species of plasmid may be exist in
these isolates.
These bacterial isolates of the P. mirabilis and P. vulgaris were chosen
for local determination of their genes that encoding resistance to 10
antibiotics and 4 heavy metals, swarming phenomenon, production of
urease, phenylalanine deaminase and production of hydrogen sulfate gas,
by the genetic transformation of the laboratory strain E. coli JM83 using
calcium chloride and heat shock. The results showed that genes confer
resistance to antiobiotics (AP, N.A, Tc, Tri, Vef, Cip, Cm, Nit, Nor& G)
and heavy metals (CoCl2, CdCl2, HgCl2& NiCl2) are located on plasmid
DNA of these chosen bacterial isolated. Also, the results of
transformation indicate that some E.coli JM83 transformant have ability
to display the swarming phenomenon and production of urease and
phenylalanine deaminase but no transformant colony can produce of H2S.
Curing of plasmid DNA content of the bacterial isolates that used in
genetic transformation experiments were performed by using the curing
agents SDS, urea and Tris with different concentrations ranging from
0.25-1.0% and at logaritmic and stationary phase of bacterial growth.
Reasonable ratio values of curing were obtained, which support the
results of genetic transformation. But the cured bacterial isolates revealed
urease, phenylalamine deaminase and H2S productio