Fifteen local isolates of Pseudomonas were obtained from several sources such as soil, water and some high-fat foods (Meat, olives, coconuts, etc.). The ability of isolates to produce lipase was measured by the size of clear zone on Tween 20 solid medium and by measuring the enzymatic activity and specific activity. Isolate M3 (as named in this study) was found to be the most efficient for the production of the lipase with enzymatic activity reached 56.6 U/ml and specific activity of 305.94 U/mg. This isolate was identified through genetic analysis of the 16S rRNA gene. and it was shown that the isolate M3 belongs to Pseudomonas aeruginosa with 99% similarity. The DNA of isolate M3 was extracted and lipase gene was amplified through PCR technique, then purified and cloned into E.coli DH5α cells first using pTG19-T plasmid, and expressed in E.coli Bl21 with expression vector pet-28a. The activity of lipase from transformed E.coli Bl21 was 196.6 U/ml and the specific activity 618.2 U/mg.
Pseudomonas aeruginosa has variety of virulence factors that contribute to its pathogenicity. Therefore, rapid detection with high accuracy and specificity is very important in the control of this pathogenic bacterium. To evaluate the accuracy and specificity of Polymerase Chain Reaction (PCR) assay, ETA and gyrB genes were targeted to detect pathogenic strains of P. aeruginosa. Seventy swab samples were taken from patients with infected wounds and burns in two hospitals in Erbil and Koya cities in Iraq. The isolates were traditionally identified using phenotypic methods, and DNA was extracted from the positive samples, to apply PCR using the species specific primers targeting ETA, the gene encoding for exotoxin A, and gyrB gene. The res
... Show MoreSTAG proteins, which are part of the cohesin complex and encoded by the STAG genes, are known as Irr1/Scc3 in yeast and as SA/STAG/stromalin in mammals. There are more variants as there are alternate splice sites, maybe three open reading frames (ORFs) code for three main proteins, including: SA1 (STAG1), SA2 (STAG2) and SA3 (STAG3). The cohesin protein complex has various essential roles in eukaryotic cell biology. This study compared the expression of the STAG1 gene in four different breast cancer cell lines, including: MCF-7, T-47D, MDA-MB-468, and MDA-MB-231 and normal breast tissue. RNA was extracted from these cell lines and mRNA was converted to cDNA, and then expression of the STAG1 gene was quantified by three sets of specific prim
... Show MoreThe 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 pJ
... Show MoreOwing to high antibacterial resistance of Pseudomonas aeruginosa, it could be considered as the main reason behind the nosocomial infections. P. aeruginosa has a well-known biofilm forming ability. The expression of polysaccharide encoding locus (pelA gene) by P. aeruginosa is essential for this ability. The purpose of the current research was to determine the biofilm formation in P. aeruginosa isolated from clinical samples and to evaluate the role of the selected PelA gene in biofilm formation using PCR method in Iraqi patients. Results revealed that 24 (96%) isolates were found to have the ability to form biofilm that was remarkably related to gentamicin resistance. Moreover, the pelA gene was found in all biofilm-producers. In c
... Show MoreIt was aimed to understand the interleukin-4 (IL-4) role in etio-pathogenesis of rheumatoid arthritis (RA). Two approaches were adopted. In the first one, a quantitative expression of IL4 gene was assessed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and such findings were correlated with some demographic, clinical and laboratory parameters, which included gender, duration of disease, disease activity score (DAS-28), rheumatoid factors (RFs), C-reactive protein (CRP) and anti-cyclic citrullinated peptide (ACCP) antibodies. In the second approach, a single nucleotide polymorphism (SNP) of IL4 gene (rs2243250) was inspected by DNA sequencing using specific primers. Fifty-one Iraqi RA patients (22 males and 29 fem
... Show MoreThe expression of the Proprotein Convertase Subtilisin/Kexin Type 9 gene (PCSK9) is inextricably related to lipid levels and a risk of atherosclerotic coronary artery disease (ASCAD). The present study aims to measure the quantity of PCSK9 gene expression and the effect of methylation on its expression level taking part in the pathogenesis of acute coronary artery disorder.
A current study included 150 subjects from the Iraqi population, 100 ASCAD patients and 50 healthy controls. The concentration of PCSK9 in each serum sample was determined by the ELISA technique, the expression levels of the PCSK9 gene in whole blood were estimated by RT-qPCR – Quantitative Reverse Transcription PCR method, and DNA
... Show MoreThe current study includes 144 samples were 106 bacterial samples belonging to the clinical sources, 38 bacterial samples belonging to the environmental sources to investigate the presence of bacteria P. aeruginosa. The results of diagnosis clarified that there are 45 bacterial isolates belonging to the bacterium P. aeruginosa The examination of the sensitivity of all bacterial isolates was done for elected 45 isolation towards the 11 antibiotic by spread method on the dishes. The results showed that the resistance ratio toward Cefixim, Cefotaxim, Tetracycline, Amoxicillin, Cloxacillin, Methicillin, Erythromycin and Naldixic acid was 77.7, 73.3, 84.4, 82.2, 80, 77.7, 77.7 and 73.3 respectively, While most isolates were sensitive to all o
... Show MoreThe density-based spatial clustering for applications with noise (DBSCAN) is one of the most popular applications of clustering in data mining, and it is used to identify useful patterns and interesting distributions in the underlying data. Aggregation methods for classifying nonlinear aggregated data. In particular, DNA methylations, gene expression. That show the differentially skewed by distance sites and grouped nonlinearly by cancer daisies and the change Situations for gene excretion on it. Under these conditions, DBSCAN is expected to have a desirable clustering feature i that can be used to show the results of the changes. This research reviews the DBSCAN and compares its performance with other algorithms, such as the tradit
... Show MoreOver the past few decades, the health benefits are under threat as many commonly used antibiotics have become less and less effective against certain illnesses not only because many of them produce toxic reactions but also due to the emergence of drug-resistant bacteria. The clinical use of a combination of antibiotic therapy for Pseudomonas aeruginosa infections is probably more effective than monotherapy. The present study aims to estimate the antibacterial and antibiofilm activity of Conocarpus erectus leaves extracts against multi-drug resistant P. aeruginosa isolated from different hospitals in Baghdad city. One hundred fifty different clinical specimens were collected from patients from September 2021 to January 2022. All samples were
... Show MoreOne of the most important virulence factors in Pseudomonas aeruginosa is biofilm formation, as it works as a barrier for entering antibiotics into the bacterial cell. Different environmental and nutritional conditions were used to optimize biofilm formation using microtitre plate assay by P. aeruginosa. The low nutrient level of the medium represented by tryptic soy broth (TSB) was better in biofilm formation than the high nutrient level of the medium with Luria Broth (LB). The optimized condition for biofilm production at room temperature (25 °C) is better than at host temperature (37 °C). Moreover, the staining with 0.1% crystal violet and reading the biofilm with wavelength 360 are considered essential factors in
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