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: It's believed that HBD-3 is involved in the tissue remodeling process of articular cartilage. Also, HBD-3 has anti-inflammatory properties. Objectives: The purpose of this study is to assay human beta-defensine-3 (HBD-3) in serum from rheumatoid arthritis (RA) patients and investigate its correlation with proinflammatory cytokines. Methods: In this case-control study, fifty-eight RA patients were aged 20–65 years, and 29 age-matched healthy subjects (HS) had no inflammatory rheumatic diseases. The disease activity score-28 joint erythrocyte sedimentation rate (DAS28-ESR) was used to measure RA activity. CRP, ACPA, HBD-3, TNF-α, and IL-1β were assessed using the enzyme-linked immunosorbent assay technique (ELISA). Res
... Show MoreThis study confirms the ubiquitin conjugating enzyme 2B (Rad6) plays a significant role in the DNA repair pathway also because the ubiquitin-conjugating pathway. The DNA repair pathway could be a variety of bypass repair mechanism where the broken base pair is bypassed by permitting the replication fork to labor under the site of injury. This is often done by a shift mechanism wherever deoxyribonucleic acid enzyme - δ is switched with DNA enzyme - η (DNAP - η). Site of DNAP - η is massive enough to permit the broken ester to labor under, and so bypass the broken nucleotide. However, this is often potential solely through the involvement of Proliferating cell nuclear antigen (PCNA) that could be a processivity issue and it acts as a plat
... Show MoreBackground: Beta thalassemia major (β-TM) is an inheritable condition with many complications, especially in children. The blood-borne viral infection was proposed as a risk factor due to the recurrent blood transfusion regimen (hemotherapy) as human parvovirus B19 (B19V). Objective: This study investigated the B19V seroprevalence, DNA presence, B19V viral load, and B19V genotypes in β-TM patients and blood donors. Methods: This is a cross-sectional study incorporating 180 subjects, segregated into three distinct groups each of 60 patients, namely control, β-TM, and β-TM infected with Hepatitis C Virus (HCV). For the B19V prevalence in the studied group, the ELISA technique and real-time PCR were used. The genotyping was follo
... Show MoreSamuel Beckett’s Happy Days (1961) clearly portrays a lack of communication among the characters of the play which refers to the condition of modern man. This failure of communication led Samuel Beckett to use a lot of pauses and silences in all plays written instead of using words. To express the bewilderment of the modern man during the 20th century, Beckett adopts the use of no language strategy in the dramatic works. After World War II, people were without hope, religion, food, jobs, homes, or even countries. Beckett gave them a voice. He used a dramatic language out of everyday things, in which silence was part of the syntax as a poetic repetition. Language is no more important to the modern man; instead, he us
... Show MoreChitinase-3-like 1 protein (YKL-40) is a glycoprotein primarily produced in the arthritic joint and plays a crucial role in inflammatory processes. The aim of the study is to establish the role of YKL-40 as a biomarker for rheumatoid arthritis (RA) compared to proinflammatory biomarkers and disease activity. The study included 58 patients and 18 control. Diseases activity score (DAS-28) and clinical disease activity index (CDAI) were measured. Serum level of YKL-40, tumor necrosis factor-α (TNF-α), interleukin-1B (IL-1β), erythrocyte sedimentation (ESR), rheumatoid factor (RF), C-reactive protein (CRP), and anti-citrullinated protein antibody (ACPA) were assessed. The results showed that the median serum YKL-40 level which was 5.42
... Show MoreIn this research, the performance of a two kind of membrane was examined to recovering the nutrients (protein and lactose) from the whey produced by the soft cheese industry in the General Company for Food Products inAbo-ghraab.Wheyare treated in two stages, the first including press whey into micron filter made of poly vinylidene difluoride (PVDF) standard plate type 800 kilo dalton, The membrane separates the whey to permeate which represent is the main nutrients and to remove the fat and microorganisms.The second stage is to isolate the protein by using ultra filter made of polyethylsulphone(PES)type plate with a measurement of 10,60 kilo dalton and the recovery of lactose in the form of permeate.
The results showed that the percen
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
... 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 MoreAbstract The present study was Conducted to evaluate the effect of amixture of three species of arbuscular mycorrhizal fungi ( Glomus etunicatum , G. leptotichum and Rhizophagus intraradices ) in Influence on the percentage of the components of NPK and protein of tomato leaves and roots infected with Fusarium oxysporum f.sp. Lycopersici wich cause Fusarial wilt disease , planted for 8 weeks in the presence of the organic matter ( peatmose) , using pot cultures in aplastic green house , Results indicated significant increase in the percentage of the elements of NK and protein of tomato leaves and roots In the control treatment (C), While the percentage of the element P was after infection with the pathogen 4 weaks after mycorrhiza
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