The bile salt hydrolase gene (bshA), encoding bile salt hydrolase enzyme (EC 3.5.1.24) from probiotic isolate Lactobacillus acidophilus Ar strain which is responsible for assimilation cholesterol were studied in the present work. About 801 bp in length DNA fragment of Lb. acidophilus Ar strain was amplified by PCR techniques. Two restriction sites (PstI/SacI) were added to each end of that fragment for manipulation of DNA during cloning. Amplified fragment inserted into pJET1.2\blunt end vector and pMG36e vector respectively. pJET1.2\blunt end vector is overexpression plasmid for E. coli MC1022, and pMG36e vector is a shuttle vector which is able to replicate in both E. coli and lactic acid bacteria. The resulted constructs were named as pJET/bshA and pMG36e/bshA respectively. Both recombinants were transferred to E. coli MC1022 by chemical transformation. Obtained recombinants analyzed for expression and sequences. The results were confirmed that production of bile salt hydrolase from recombinant E. coli MC1022 pJET/bshA found to be higher while compared with E. coli MC1022 wild type and recombinant E. coli MC1022 pMG36e/bshA strain. However production of bile salt hydrolase from recombinant E. coli MC1022 pMG36e/bshA found to be higher while compared with E. coli MC1022 wild type. The recombinant plasmid also found to be stable in host organism after a few generations.
Inhibitory effect of Lactobacillus plantarum was studied against Escherichia coli O157:H7 in minced beef , as well as the effect of some Spices ( Garlic , Cloves and Black Pepper ) was studied against this bacteria . Plate count method was used to estimate the viable cells of E. coli O157:H7 in the treated meat samples for incubation periods( 0 , 24 and 48) h. Results showed that Lactobacillus plantarum had inhibitory effect against E. coli O157:H7 , when reducing the number of cells after 24, 48 h., and reached to Zero after (24) h. As well as Spices in this study had inhibitory activity against E. coli O157:H7 .Garlic had
... Show MoreThe present study aims to detect CTX-M-type ESBL from Escherichia coli clinical isolates and to analyze their antibotic susceptibility patterns. One hundred of E. coli isolates were collected from different clinical samples from a tertiary hospital. ESBL positivity was determined by the disk diffusion method. PCR used for amplification of CTX-M-type ESBL produced by E. coli. Out of 100 E. coli isolates, twenty-four isolates (24%) were ESBL-producers. E. coli isolated from pus was the most frequent clinical specimen that produced ESBL (41.66%) followed by urine (34.21%), respiratory (22.23%), and blood (19.05%). After PCR amplification of these 24 isolates, 10 (41.66%) isolates were found to possess CTX-M genes. The CTX-M type ESBL
... Show MoreThis study was aimed to detect weather copA gene(copper resistance gene ) presence in A.baumannii A92 genome(AbaR genomic islands). The full genomic sequence of A.baumannii A92 not published in NCBI genome similarity was detected between two strains so the sequence of A.baumanniiIS-116(Genebank-AMGF0100000.1 ) was used to design the primers that were used for amplify of copA gene of A.baumannii A92.
Two primers contain two sites for restriction enzymes (KpnI,XohI) and PWSK29 vector were used in the cloning, double digestion has been performed for vector and gene. Then the re-ligation was completed to form recombinant molecule,after that, transformation have been performed for the recombinant molecule by using chemical competent E.coli
Four hundred and fifty urine samples were collected from patients suffering from urinary tract infection from the General Azadi hospital in Kurkuk province ,during the period of october 2007 till march 2008 . Results of bacteriological culture revealed that (168) out of (450) studied samples (37.3%) gave positive culture using blood agar and macConkey agar ; different species of bacterial isolates were detected using morphological and biochemical tests ,from these isolates the highest percentage of the isolates were from Escherichia coli when it was (100) isolates out of (190) isolate (52.63%) . one hundred isolates were distributed between (77) from females and (23) from
... Show MoreThe effect of crude colicin extracted from E. coli isolated from
urinary tract infection patients on phagocytosis in vitro was studied . Results showed that the effect of crude colicin on phagocytic cells and their activities were concentration dependent. Low concentration (50
Mg/ml) have no significant effect (p>O.OS) on shape, size, migration and engulfment activity of phagocytic cells, while (50, 100 Mg/ml)
enhanced killing activity and increase superoxide ( 02 ) production as indicated by (NBT) test. But high concentration (500 mg/ml) of crude colicin caused inhibition of phagocytic cel
... Show MoreA significant increase in the incidence of non-O157 verotoxigenic Escherichia coli (VTEC) infections have become a serious health issues, and this situation is worsening due to the dissemination of plasmid mediated multidrug-resistant microorganisms worldwide. This study aims to investigate the presence of plasmid-mediated verotoxin gene in non-O157 E. coli. Standard microbiological techniques identified a total of 137 E. coli isolates. The plasmid was detected by Perfectprep Plasmid Mini preparation kit. These isolates were subjected to disk diffusion assay, and plasmid curing with ethidium bromide treatment. The plasmid containing isolates were subjected to a polymerase chain reaction (PCR) for investigating
... Show MoreThirty uropathogenic E. coli isolates were isolated from hospitalized and non hospitalized patients, complaining of urinary tract infections, of Al-Kadhymia Teaching Hospital and subjected to tRNA extraction. A method of tRNA extraction was modified by adding sodium dodecyl sulfate (SDS) instead of urea. Polyacrylamide gel electrophoresis and two methods of staining, ethidium bromide staining and silver staining, as well as spectrophotometric detection were used.
Background: The beneficial gut bacterium E. coli can cause blood poisoning, diarrhoea, and other gastrointestinal and systemic disorders. Objective: This study amid to examines the antibiofilm activity of Laurus nobilis leaves extract on E. coli isolates and compares pre- and post-treatment gene expression of fimA and papC genes. Subjects and Methods: Ten isolates of E. coli were obtained from the Genetic Engineering and Biotechnology Institute, University of Baghdad, which was previously collected from Baghdad city hospitals and diagnosed by chemical tests, the diagnosis was confirmed using VITEK-2 System. The preparation of the aqueous and methanolic Laurus nobilis leaves extracts was done by using the maceration method and Soxhlet appara
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