Expression of thermostable MMLV reverse transcriptase in Escherichia coli by directed mutation.

The functionality of Moloney murine leukemia virus reverse transcriptase (MMLV RT) will increase with the improvement of its solubility and thermal stability. Introduce directed mutation at specific positions of the MMLV RT sequence and codon optimization is needed to achieve these properties. The two RT coding sequences with (rRT-K) and without directed mutations (rRT-L) were versatility optimized and expressed to analyze the ribonuclease H (RNase H) inactivity and thermostable polymerase activity.

RNA-Seq transcriptome profiling of immature grain wheat is a technique for understanding comparative modeling of baking quality.

Improving the baking quality is a primary challenge in the wheat flour production value chain, as baking quality represents a crucial factor in determining its overall value. In the present study, we conducted a comparative RNA-Seq analysis on the high baking quality mutant "O-64.1.

Augmenting the Antitumor Efficacy of Natural Killer Cells via SynNotch Receptor Engineering for Targeted IL-12 Secretion.

Natural killer (NK) cells are crucial components of innate immunity, known for their potent tumor surveillance abilities. Chimeric antigen receptors (CARs) have shown promise in cancer targeting, but optimizing CAR designs for NK cell functionality remains challenging. CAR-NK cells have gained attention for their potential to reduce side effects and enable scalable production in cancer immunotherapy.

Effects of Bacillus in Pectobacterium quorum quenching: A survey of two different acyl-homoserine lactonases.

Numerous functions in pathogenic Pectobacterium are regulated by quorum sensing (QS). Two different aiiA genes isolated from Bacillus sp. A24(aiiA) and Bacillus sp.

Evaluation of the impact of polypeptide-p on diabetic rats upon its cloning, expression, and secretion in Saccharomyces boulardii.

This study was designed to evaluate the effectiveness of recombinant polypeptide-p derived from Momordica charantia on diabetic rats. In this research, the optimized sequence of polypeptide-p gene fused to a secretion signal tag was cloned into the expression vector and transformed into probiotic Saccharomyces boulardii. The production of recombinant secretion protein was verified by western blotting, HPLC, and mass spectrometry.

Enhancing productivity of Chinese hamster ovary (CHO) cells: synergistic strategies combining low-temperature culture and mTORC1 signaling engineering.

The growing demand for recombinant proteins in medicine has prompted biopharmaceutical companies to seek ways to maximize the manufacturing process. Despite its known negative impact on cell growth, temperature shift (TS) has emerged as a cost-effective strategy to enhance protein quantity and quality in Chinese Hamster Ovary cells (CHO). As cells adapt their growth and protein synthesis rate to the environment through influencing mTOR complex 1 (mTORC1), here we evaluated the potential of mTORC1 signaling engineering to improve the production of granulocyte-macrophage colony-stimulating factor (GM-CSF) protein in stable CHO cells at low temperature.

Designing a Multi-epitope Vaccine against the SARS-CoV-2 Variant based on an Immunoinformatics Approach.

Background: SARS-CoV-2 is a life-threatening virus in the world. Scientific evidence indicates that this pathogen will emerge again in the future. Although the current vaccines have a pivotal role in the control of this pathogen, the emergence of new variants has a negative impact on their effectiveness.

A simple and efficient method for cytoplasmic production of human enterokinase light chain in E. coli.

Human enterokinase light chain (hEKL) cDNA sequence was designed with the help of codon optimization towards Escherichia coli codon preference and ribosome binding site design and artificially synthesized with a thioredoxin fusion tag at the N-terminal and a five his-tag peptide at the C-terminal. The synthetic hEKL gene was cloned into the pET-15 expression vector and transferred into the three different expression strains of E. coli BL21(DE3), NiCo21, and SHuffle T7 Express.

RT-LAMP in SARS-CoV-2 detection: point to improve primer designing and decrease molecular diagnosis pitfalls.

Background: Due to the high transmission rate of SARS-CoV-2, diagnostic tests have become tools for identifying patients. The key points were the virus genomes survey to design RT-LAMP primers; comparing the sensitivity and specificity of RT-LAMP and RT-qPCR; and determining the relationship among clinical symptoms, CT scan, RT-qPCR, and RT-LAMP results.

Methods: This cohort study included 444 symptomatic patients.

Curcumin inhibits the replication of rotavirus in vitro.

Rotavirus is the most important etiological agent of infectious diarrhea in children under 5 years of age with more than 125,000 deaths occurring annually worldwide. The present study aims to determine the effect of curcumin, a natural polyphenol compound, on rotavirus in a cell culture model. The anti-viral activity of curcumin was evaluated by reverse-transcriptase quantitative PCR (RT-qPCR), TCID50, and western blot techniques to assess CC50 in curcumin-treated MA104 cells as well as EC50 and SI within the infected MA104 cell line.

Ion content, antioxidant enzyme activity and transcriptional response under salt stress and recovery condition in the halophyte grass Aeluropus littoralis.

Objective: In contrast to glycophytes, halophyte plants have evolved unique morphological and physiological mechanisms to deal with abiotic stress. This study presents the physiological responses of Aeluropus littoralis, a halophyte grass, to salt stress and recovery conditions on the molecular level.

Results: Elemental analysis showed that Na concentration increased in the analyzed tissue during salt stress application, and declined at recovery condition.

Identification of salinity responsive genes in lavender through cDNA-AFLP.

Currently, a global demand exists forlavender as a significant medicinal plant and source of essential oils. Freshwater and arable lands are two major factors that inhibit extensive farming of medicinal plants in Iran. Saline water from seas and salty soil may be new resources for agricultural use, especially for medicinal plants.

The effect of pomegranate (Punica granatum) supplementation on metabolic status in patients with type 2 diabetes: A systematic review and meta-analysis.

Objective: Several randomized controlled clinical trials (RCTs) have investigated the effect of pomegranate (Punica granatum) supplementation in patients with type 2 diabetes mellitus (T2DM), with conflicting results. Therefore, the present study assessed the effect of pomegranate on metabolic status and oxidative stress in patients with T2DM.

Methods: A comprehensive search was conducted in Pubmed, EMBASE, ISI web of science, Cochrane library and Scopus databases until April 2020, to identify RCTs investigating the effect of pomegranate supplementation on glycemic markers (fasting blood glucose (FBG), homeostasis model assessment-estimated insulin resistance index (HOMA-IR), glycosylated hemoglobin percentage (HbA1c) and insulin levels), anthropometric indices (body mass index (BMI) and body weight (BW)), blood lipids (triglycerides (TG), total cholesterol (TC), high lipoprotein cholesterol (HDL-C) and low lipoprotein cholesterol (LDL-C)), inflammatory markers (high sensitive c-reactive protein (Hs-CRP)) and markers of oxidative stress (total antioxidant capacity (TAC)) in adults with T2DM.

Advanced sequence optimization for the high efficient yield of human group A rotavirus VP6 recombinant protein in Escherichia coli and its use as immunogen.

Rotavirus is the important etiological agents of infectious diarrhea among children under 5 years old. Rotaviruses are divided into 10 serogroups (A-J) and each group is based on genetic properties of major structural protein VP6. We designed a novel VP6 sequence optimization to increase the expression level of this protein.

Identification of hub genes associated with RNAi-induced silencing of XIAP through targeted proteomics approach in MCF7 cells.

Background: The X-linked inhibitor of apoptosis protein (XIAP) is the most potent caspase inhibitor of the IAP family in apoptosis pathway. This study aims to identify the molecular targets of XIAP in human breast cancer cells exposed to XIAP siRNA by proteomics screening. The expression of XIAP was reduced in MCF-7 breast cancer cells by siRNA.

Lead-induced oxidative stress and role of antioxidant defense in wheat ( L.).

The aim of this study was to investigate soil lead pollution on biochemical properties and gene expression pattern of antioxidant enzymes in three wheat cultivars (Morvarid, Gonbad and Tirgan) at flag leaf sheath swollen stage. Lead (Pb(NO)) was used at four different concentrations (0, 15, 30 and 45 mg/kg of soil). The leaf and roots samples were taken at late-booting stage (Zadoks code, GS: 45).

Proteomics evaluation of MDA-MB-231 breast cancer cells in response to RNAi-induced silencing of hPTTG.

Aims: Breast cancer is the most common cancer in women worldwide. Several genes are up-regulated in breast cancer such as human pituitary tumor transforming gene (hPTTG). This study aims to evaluate cell proliferation and the downstream expression pattern of hPTTG1 gene at the mRNA and protein levels after specific down-regulation of hPTTG1 by siRNA.

New insights into the roles of the and genes in signaling pathways.

The forkhead box O3 (FOXO3) and p27Kip1 are two important genes in breast cancer progression. In the present study we analyzed the effect of simultaneous FOXO3 silencing and p27Kip1 activation on breast cancer cell survival and the potential targets of these changes in cancer molecular pathways. The present study involved the cloning of shRNA and genes under the control of the bidirectional survivin promoter to down- and up-regulate and genes, respectively.

Differential molecular response of maize and Johnson grass against maize dwarf mosaic virus and bermuda grass southern mosaic virus.

Maize dwarf mosaic virus (MDMV) and bermuda grass southern mosaic virus (BgSMV) is the most important cereal potyvirus in Iran. Expression of some key genes in maize plants susceptible and tolerant to MDMV or BgSMV and gene expression profile of MDMV and BgSMV compatible or incompatible to Johnson grass plants were studied. Time points of 1, 9, 24 and 72 h after inoculation with both viruses were investigated as well.

CDC25A pathway toward tumorigenesis: Molecular targets of CDC25A in cell-cycle regulation.

The cell division cycle 25 (CDC25) phosphatases regulate key transitions between cell-cycle phases during normal cell division, and in the case of DNA damage, they are key targets of the checkpoint machinery that ensure genetic stability. Little is known about the mechanisms underlying dysregulation and downstream targets of CDC25. To understand these mechanisms, we silenced the CDC25A gene in breast cancer cell line MDA-MB-231 and studied downstream targets of CDC25A gene.

RNA-Seq analysis revealed genes associated with drought stress response in kabuli chickpea (Cicer arietinum L.).

Drought is the most important constraint that effects chickpea production globally. RNA-Seq has great potential to dissect the molecular mechanisms of tolerance to environmental stresses. Transcriptome profiles in roots and shoots of two contrasting Iranian kabuli chickpea genotypes (Bivanij and Hashem) were investigated under water-limited conditions at early flowering stage using RNA-Seq approach.

Assessment of DNA Contamination in RNA Samples Based on Ribosomal DNA.

One method extensively used for the quantification of gene expression changes and transcript abundances is reverse-transcription quantitative real-time PCR (RT-qPCR). It provides accurate, sensitive, reliable, and reproducible results. Several factors can affect the sensitivity and specificity of RT-qPCR.

A novel purple acid phytase from an earthworm cast bacterium.

Background: Phytases are a diverse group of enzymes initiating the dephosphorylation of phytate. Phytate is considered as an anti-nutritional compound because of its capability to chelate nutrients such as Fe , Zn , Mg , and Ca . In this study, several bacterial isolates obtained from earthworm casts were evaluated for their phytate degrading capability.

Proteomics analysis of Medicago truncatula response to infection by the phytopathogenic bacterium Ralstonia solanacearum points to jasmonate and salicylate defence pathways.

The infection of the model legume Medicago truncatula with Ralstonia solanacearum GMI1000 gives rise to bacterial wilt disease via colonisation of roots. The root and leaf responses to early infection (1 and 3 days post infection) were characterised to investigate the molecular mechanisms of plant resistance or susceptibility. A proteomics approach based on pools of susceptible and resistant recombinant inbred lines was used to specifically target the mechanisms for tolerance.

Study on the chemical and microbial composition and probiotic characteristics of dominant lactic acid bacteria in fermented poultry slaughterhouse waste.

This study aimed to evaluate the quality of fermented poultry slaughterhouse waste and isolate and identify the probiotic bacteria present in it. Poultry slaughterhouse waste samples supplemented with 5% starter culture and 0.2% propionic acid received one of four experimental treatments, which consisted of further supplementation with: (1) 10% molasses (M10); (2) 15% molasses (M15); (3) 10% molasses with 200ppm butylated hydroxytoluene (MB10); and (4) 15% molasses with 200ppm butylated hydroxytoluene (MB15).