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Reliable Reference Gene for Normalization of RT- qPCR Data in Human Cancer Cell Lines
Subjected to Gene Knockdown
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Quantitative real-time Polymerase Chain Reaction (RT-qPCR) has become a valuable molecular technique in biomedical research. The selection of suitable endogenous reference genes is necessary for normalization of target gene expression in RT-qPCR experiments. The aim of this study was to determine the suitability of each 18S rRNA and ACTB as internal control genes for normalization of RT-qPCR data in some human cell lines transfected with small interfering RNA (siRNA). Four cancer cell lines including MCF-7, T47D, MDA-MB-231 and Hela cells along with HEK293 representing an embryonic cell line were depleted of E2F6 using siRNA specific for E2F6 compared to negative control cells, which were transfected with siRNA not specific for any gene. Using RT-qPCR, Ct (threshold cycle) values of 18S and ACTB were determined in transfected cells and compared with control cells. In the selection of the above cell lines, 18S was identified as the most stably expressed reference gene than ACTB in gene knockdown experiments.

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Publication Date
Sun Dec 26 2021
Journal Name
Sultan Qaboos University Journal For Science [squjs]
The Deubiquitylase USP5 Knockdown Reduces Semliki Forest Virus Replication in HeLa Cells
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Semliki Forest Virus (SFV), a member of the Alphavirus genus in the Togaviridae family, is a small-enveloped, positive-sense single-stranded RNA (+ssRNA) virus. The virus is spread by mosquitos and can infect humans, resulting in mild febrile disease with symptoms that include fever, myalgia, arthralgia, persistent headaches and asthenia.  Virulent strains of SFV in mice cause lethal encephalitis by infecting neurons in the central nervous system. In on-going experiments in the research group using a focused siRNA screen we have investigated the role of deubiquitylases (DUBs) during SFV infection (as a model alphavirus) and monitored the effect of DUB depletion on cell viability after infection. We identified a group of DUBs that h

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Publication Date
Mon Jan 01 2024
Journal Name
Baghdad Science Journal
Molecular Identification of Methylorubrum extorquens using PCR-Amplified MxaF Gene Fragments as A Molecular Marker
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  Methylotrophs bacteria are ubiquitous, and they have the ability to consume single carbon (C1) which makes them biological conversion machines. It is the first study to find facultative methylotrophic bacteria in contaminated soils in Iraq. Conventional PCR was employed to amplify MxaF that encodes methanol dehydrogenase enzyme. DNA templates were extracted from bacteria isolated from five contaminated sites in Basra. The gene specific PCR detected Methylorubrum extorquens as the most dominant species in these environments. The ability of M. extorquens to degrade aliphatic hydrocarbons compound was tested at the laboratory. Within 7 days, gas chromatographic (GC) studies of remaining utilize

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Publication Date
Mon Apr 01 2019
Journal Name
Biochemical And Cellular Archives
Association Of Human Cytomegalovirus With Her2 Protooncogene Overexpression In Iraqi Breast Cancer Patients
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Human cytomegalovirus (HCMV) has a worldwide distribution and common infections. The presence of HCMV genome and antigens has been detected in many kinds of human cancers. The proto-oncogene Her2/neu is overexpressed in 25% to 30% of human breast cancers and frequently associated with tumor aggressiveness and worse prognosis. Examination was applied to detect different HCMV antigens and to demonstrate their correlation with Her2/neu overexpression in breast cancer. The present study includes samples from 70 women of 60 breast cancer patients and 10 normal breast tissues. Formalin-fixed paraffin embedded tissue blocks were obtained from each woman according to ethical approval. Human cytomegalovirus early antigen expression was detected in 5

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Publication Date
Tue Jan 01 2019
Journal Name
Iraqi Journal Of Agricultural Sciences
Cloning and expression of a lipase gene from Pseudomonas aeruginosa into E.coli
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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 tec

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Publication Date
Thu Oct 01 2020
Journal Name
Biochemical & Cellular Archives
STUDY ON ABILITY OF PROTEUS MIRABILIS ISOLATED FROM BURNS AND WOUNDS INFECTIONS TO BIOFILM FORMATION AND DETECTION URE C GENE RESPONSIBLE FOR PRODUCE UREASE ENZYME
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A total of 200 clinical samples included Burns and Wounds infections were collected from Baghdad Governorate. Results showed that rate all isolates of P. mirabilis was 31(15.5%) and rate of Burns infections was 14 (45%) and rate of wounds infection 17 (55%). Where was diagnostic based on conventional biochemical tests and confirmed by the Vitek-2 Compact system and the specific primer of the16SrRNA gene, the ability of bacterial isolates to biofilm formation to be studied. It's considered an important virulence factor in Incidence of diseases and play important role in increasing resistance to antibiotic of encased bacteria, by two methods Congo Red Agar method and Microtiter Plate method. The Congo Red Agar method showed that most isolates

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Publication Date
Sun May 10 2020
Journal Name
Baghdad Science Journal
Lack of Association between LCS6 Variant in KRAS Gene with the Occurrence of Breast Tumors in Iraqi Women
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Breast cancer is the most commonly diagnosed cancer and remains one of the main reasons of cancer-related mortality in women worldwide. KRAS variant rs61764370 (T>G) is associated with an increased risk of occurrence of many cancers, Here The case-control study was accomplished on 135 women including 45 women with breast cancer patients, 45 women with benign breast lesions and 45 healthy women to analyze the association of KRAS variant rs (61764370 T>G) with breast cancer. LCS 6 variant in KRAS gene was amplified by using specific primers, then genotype was detected after sequencing the PCR products. The results showed that the genotype and allele frequency of TT and GT allele of  KRAS

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Publication Date
Tue Mar 15 2022
Journal Name
Journal Of Baghdad College Of Dentistry
In vitro cytotoxic effect of annona squamosa pulp ex-tract as a mouthwash for children on human normal cell line
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Background: Fruits and their by-products are the primary sources of bioactive chemicals in plants. Because of its phytochemical richness, Annona squamosa fruits have gained the alertness of people willing in health-promoting diets. The purpose of this in vitro study was to evaluate the cytocompatibility effect of ethanolic crude extract of Annona squamosa pulp against a human normal cell line as a mouthwash for children. Material and method: The ethanolic extract of Annona squamosa pulp was extracted using the ultrasonic method and then lyophilized to make it powder. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) test was performed to investigate the cytotoxic activity of the pulp extract on a human normal cell l

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Publication Date
Tue Jan 01 2019
Journal Name
Kirkuk University Journal
DNA Sequences of qacE∆1 gene in Pseudomonas aeruginosa isolated from wounds and burns infections.
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Publication Date
Wed Apr 01 2020
Journal Name
Malaysian Journal Of Biochemistry And Molecular Biology
Mutations in ergosterol 11 gene of fluconazol resistant candida albicans isolated from different clinical samples
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Fluconazole was used to test the susceptibility of Candida albicans isolated from different clinical samples, and to detect mutations in ERG11 gene, and their relationship to fluconazole resistance. Forty-eight isolates of Candida albicans were tested for susceptibility using the disc diffusion method (M-44). ERG11 genes of six isolates were amplified (four resistant, two susceptible) and sequenced. The sequenced genes were analyzed to detect the mutations. Out of 48 isolates of Candida albicans, 4 (8%) were resistant to fluconazole. Sixteen-point mutations were detected included 13 silent mutations, and three missense mutations. The mutations of A945C (E266D) and G1609A (V488I) were found only in susceptible Candida albicans isolates, whil

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Publication Date
Sun Jan 01 2017
Journal Name
Disease Markers
Genotyping of<i> IL-4</i> −590 (C&gt;T) Gene in Iraqi Asthma Patients
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This study is the first investigation in Iraq dealing with genotyping of IL-4 −590 (C>T) gene, especially in Iraqi patients with asthma. We studied forty-eight blood samples collected from patients with asthma and compared with age-matched 25 healthy individuals as controls. The polymorphism results of IL-4 −590 (C>T) gene by using amplification refractory mutation system (ARMS-PCR) showed the presence of C and T alleles and three genotypes (CC, CT, and TT). Interestingly the frequency of C allele and CC genotype was higher in patients with asthma in comparison with the same allele and geno

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