Molecular farming has become one of the most significant implementations of modern biotechnology to generate modified plant crops to produce medicinal proteins. Agrobacterium is one plant genetic engineering tool that integrates genes of interest inside a host plant. In recent years, the need to produce recombinant proteins as therapeutics has growing rapidly, and human glucocerebrosidase is one of the proteins that is need to treat disease. In this study, specific primers were designed to amplify Hu-GBA1 gene from constructed pGEM-GBA plasmid which was cloned into the plant expression vector pCAMBIA1304. The generated recombinant pCAMBIA1304-GBA plasmid was used to transform A. tumefaciens LBA4404 and applied for transformation of sunflower cotyledon explants. Colony PCR technique was used to confirm the presence of Hu-GBA1 gene in transformed A. tumefaciens. Agrobacterium containing pCAMBIA1304-GBA was suspended in Infection Medium (IM) supplement with 200 mM acetosyringone. A bacterial suspension was used to transform sunflower cotyledons. After infection, cotyledons were co-cultivated in Co-cultivation medium (CCM), supplied with 200 mM acetosyringone without antibiotics. The cotyledons were then transferred to selection media containing 7.5 mg/L Hygromycin and 250 mg/L Cefotaxime and grown for additional 14 days at 25℃ in photoperiod of 16h L/8h D. The transformed sunflower cotyledons were successfully generated complete plant with used 6-Benzylaminopurine and Naphthalene acetic acid as growth hormones. The presence of the Hu-GBA1 gene in the genomic DNA of transgenic sunflower plant was proven by PCR as a band of 1561bp size. The GBA mRNA expression in modified sunflowers was detected by qRT-PCR compared with control GBA mRNA. Enzyme Linked Immunoassay was done on crude recombinant protein that extracted from transformed sunflower using Human Glucosylceramide ELISA Kit, the Elisa test results confirmed the production of recombinant glucocerebrosidase and the concentration of crude recombinant enzyme extracted from transformed sunflower with GBA1 gene was 0.45 ng/µl
Background: Colorectal Adenocarcinoma contributes one of the most common malignancies and the second leading cause of death from cancer in the western world. Epidermal growth factor receptor (EGFR) belongs to a family (ErbB-tyrosine receptors), EGFR plays an important role in the pathogenesis of colorectal cancer.
Objectives: to evaluate the immunohistochemical expression of epidermal growth factor receptors (EGFR) in colorectal adenocarcinoma and to correlate this expression with different clinicopathological parameters.
Pateints and method: In this study clinicopathological parameters of twenty five colorectal adenocarcinoma cases diagnosed in private pathology laboratories in Baghdad /Iraq from November 2012 to September 2013 we
Background:- Colorectal carcinoma is the most common cancer after the breast cancer in female and bronchus cancer in male. P53 is a tumor suppressor gene, approximately half of colorectal cancers present mutation in p53 gene.
Objectives:- To determine the frequency and the pattern of p53expression in colorectal carcinoma by immunohistochemical technique and to correlate this expression with different clinicopathological parameters.
Materials and methods:-Thirty cases of colorectal carcinoma were included in this study, these cases were diagnosed in private pathology laboratories in Baghdad / Iraq from January 2015 to Jaune 2015. Clinicopathological parameters such as age , gender , pathological diagnosis , including the tumor site
The 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 MoreBackground: Colorectal carcinoma is the most frequent type of malignancy in terms of incidence,it is the third most common cancer world wide in both sexes . bcl_2 expression has been described as been a better prognostic factor in colorectal carcinoma.
Objcetives: To determine the frequency and the pattern of Bcl-2 expression in colorectal carcinoma by immunohistochemical technique and to correlate this expression with different clinicopathological parameters.
Material and method: Thirty cases of colorectal carcinoma were studied, these cases were diagnosed in private laboratories in Baghdad / Iraq from January 2015 to Juan 2015. Clinicopathological parameter such as age, gender , pathological diagnosi
Pseudomonas aeruginosa is an opportunistic pathogen responsible for serious infections. At least three different exopolysaccharides, alginate, polysaccharide synthesis locus (Psl), and pellicle exopolysaccharide (Pel) make up the biofilm matrix in P. aeruginosa . The effect of temperature on the biofilm formation and gene expression was examined by microtiter plate and real-time quantitative polymerase chain reaction (qRT-PCR). To be able to determine the effect of temperature on biofilm formation and gene expression of P. aeruginosa, 303 clinical and environmental samples were collected. Pseudomonas aeruginosa was isolated from 61 (20.1%) and 48 (15.8%) of the clinical and e
... Show MoreCoagulation is the most important process in drinking water treatment. Alum coagulant increases the aluminum residuals, which have been linked in many studies to Alzheimer's disease. Therefore, it is very important to use it with the very optimal dose. In this paper, four sets of experiments were done to determine the relationship between raw water characteristics: turbidity, pH, alkalinity, temperature, and optimum doses of alum [ .14 O] to form a mathematical equation that could replace the need for jar test experiments. The experiments were performed under different conditions and under different seasonal circumstances. The optimal dose in every set was determined, and used to build a gene expression model (GEP). The models were co
... Show MoreThis study aimed to evaluate the IHC expression of CDX2 protein in HGC patients and control groups and also to study the correlation between IHC expression of the CDX2 and different clinicopathological variables such as: age, gender, histopathological subtype, grade, and stage of the tumor in HGC cases. the retrospectively sectional study for the period from 2014 to 2018 included a total of 60 formalin fixed paraffin embedded blocks of the HGC tissue (partial or total gastrectomy specimens) that collected from the archived materials of the Department of Pathology of Baghdad Teaching Hospital and the Center of Gastrointestinal and Hepatic Diseases, and also some samples were collected from other private laboratories. The IHC expression of th
... Show MoreFumonisin B1 (FB1) is a mycotoxin produced in some grains (mainly corn) by Fusarium species. Due to a structural similarity between FB1 and sphinganine, sphingolipids metabolism is inhibited. Such inhibition plays a critical role in cell to cell singling and structure of lipoprotein; therefore FB1 has been suggested to have a relationship with human and animal cancer. This research is planned to study the effect of FB1 on male mice at two doses (20 and 30 µg/ ml) on the expression of TGF-β1 and p16 in liver cells. Three groups of Swiss albino male mice; each group was orally administrated with FB1 toxin as the following: normal saline (control group); 20 and 30 µg/ ml. All groups were sacrificed after two weeks of oral manage
... Show MoreAntibiotic resistance is the capability of the strains to resist or protect themselves from the effects of an antibiotic. Such a resistance towards the current antimicrobials leads to the search of novel antimicrobials. Nanotechnology has been promising in different field of science and among it is the use of nanoparticles as antibacterial agents. The gastrointestinal tract seems to be the primary reservoir of uropathogenic E.coli (UPEC) in humans. UPEC strains harbour the urinary tract and cause urinary tract infection. They cause serious ailments in terms of humans. They develop resistance and increase their virulence by forming biofilms. They also show a remarkable locomotory movement with the aid of autoinducer controlled ge
... Show MoreBackground: Colorectal adenocarcinoma is the most common type of gastrointestinal cancer, with about 150,000 cases each year in the United States and about 56,730 deaths from colorectal cancer risk. In Iraq and according to the Iraqi cancer registry (ICR) reports, the incidence of colorectal carcinoma was 4.55% of whole body malignancy, & it is the seventh cause of death from cancer (ICR 2005). In males, it’s the 7th common cancer while in females it’s the 4th most common cancer. The estrogen and progesterone receptors belong to a super- family of nuclear hormone receptors. These receptor proteins function as transcription factors when they are bound to their respective ligands, these receptors shares a common st
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