Transient expression of a green fluorescent protein in tobacco and maize chloroplast

Maize is considered to be one of the staple crops across the world. However, the limited production and unforeseen weather conditions often limit its availability. This is the major reason that scientists across the globe are working towards finding ways to improve the stability of the maize crop in harsh environmental conditions and its overall production rate. The improved quality of maize will not only help meet the demand of the ever-growing global population but will also overcome the shortage due to unanticipated environmental conditions. To achieve this, researchers tried for maize plastid transformation, which is not achieved effectively yet due to the recalcitrant conditions of the crop.

In the present study, researchers constructed two vectors containing homologous recombination sequences from maize and grass. These vectors are designed to later integrate into the chloroplast genome from an inverted repeat region. The vectors consist of two crucial genes mgfp5 and hph gene (as selection marker). The former gene is driven by Prrn, a leader sequence of the atpB gene and a terminator sequence from the rbcL gene. Whereas the later is driven by Prrn, a leader sequence from rbcL gene and a terminator sequence from the rbcL gene. The vectors were then used to transform the explants of maize, tobacco, and E.coli to assess the transitory expression. The expression levels were evident from the green and red fluorescent light when observed under the epiflourescence microscope.

The results of the study show the successful expression of both vectors, along with the presence of a reporter gene in all three organisms. This highlights the capability of vectors to express genes in the cell compartments. The results in the paper are the first report of transient expression of GFP in maize embryos, offering the opportunity to improve the recalcitrant crops genetically using biotechnological interventions. 

An in situ-Synthesized Gene Chip for the Detection of Food-Borne Pathogens on Fresh-Cut Cantaloupe and Lettuce

Food-borne pathogens are one of the major reasons behind endangering the life and safety of people across the globe. Fresh foods are specifically more vulnerable to these pathogens, making it crucial to have a very efficient food safety surveillance technology. The development of such technology will help in offering rapid detection of food-borne pathogens. In the present study, researchers developed an In-situ synthesized gene chip for the detection of the food-borne pathogen. Here the researchers first identified and screened the target genes by comparing the sequences of common food-borne pathogens like Salmonella, Vibrio parahemolyticus, Staphylococcus Aureus, Listeria monocytogenes and E.coli 0157:H7 from the NCBI database. Unique tilling array probes were designed that helps to target the selected genes in an optimized hybridization system. The resultant assay showed high specificity along with strong amplification signals. The results were highly accurate with a detection limit of approximately 3 log cfu/g without culturing. The detection time for the five target food-borne pathogens on the fresh-cut cantaloupe and lettuces was found to be 24 hours. This highlights the great efficacy of the detection system to rapidly monitor the pathogens on the fresh food items. Such a system can be easily incorporated as an efficient food surveillance system for checking the logistical distribution chain, the food at the processing stage, cleaning condition at the food manufacturing plants, transport, sales and more. The technology is considered valuable as it supports the safety of fresh agricultural products, reducing the overall wastage of food due to infectious pathogens.

COVID 19: A battle between heat and humidity Vs Sunshine

Enhanced Thermostability and Anticancer Activity in Breast Cancer Cells of Laccase Immobilized on Pluronic-Stabilized Nanoparticles

Laccases are mainly found in fungi and plants known for catalyzing oxidation reactions. They are often named as multi-copper enzymes having a wide range of applications across a multitude of biotechnological processes.  Though widely used the major challenge is the low stability of the free enzyme which often restricts its usability for industrial purposes.  The development of new methods that can enhance the enzymatic activity is a need of time. In the present study, immobilization of trametes versicolor laccase on pH-responsive, pluronic stabilized silver nanoparticles (AgNPsTrp) was carried out. The results showed enhanced activity if enzyme upon stabilization of AgNPsTrp with amphiphilic copolymer Pluronic F127, this helps in changing the microenvironment of active site facilitating the improved activity. The results were confirmed using circular dichroism (CD) and fluorescence spectroscopy. Further analysis showed that lowering the activation energy and expanding the temperature window for substrate hydrolysis can play a crucial role in improving the enzyme activity. No change in the behavior of the nanocomposite indicated the stability of the enzyme as normal with the absence of any aggregation after immobilization. The efficiency of the nanocomposite in breast cancer cells- MCF-7 was observed which showed inhibition of cell proliferation by promoting cell apoptosis and beta-estradiol degradation. Further, qRT- PCR experiments were conducted to understand the underlying molecular mechanism hindering cell proliferation. The analysis showed a reduced level of mRNA levels of the anti-apoptotic genes and enhancement in level of pro-apoptotic genes. For example, BCL-2 and NF-kβ, and increase in levels of pro-apoptotic genes like p53. The entire study provides a new way of enhancing the enzymatic activity of Laccases and extending its subsequent applications.

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Cellulase and xylanase synergism in industrial biotechnology

Cellulase and Xylanase synergism in industrial biotechnology

In recent years, many studies have been carried out which shows tremendous benefits of biocatalyst when compared to chemical catalysts. Biocatalyst helps in carrying out reactions in an environment-friendly manner while providing a high level of selectivity, specificity, and low energy consumption.  Two of the most widely used enzyme as biocatalyst are cellulose and xylanase wherein the former is considered to be the 3rd highest used enzyme. Many of the industrial applications require the use of both the enzyme in combination such as bioethanol production, removal of ink in waste paper industries, food processing industries, feed processing, removal of fibers from the textile materials, pulp production, pharmaceuticals and many more. Cellulose and xylanase are mainly produced by microorganisms which increases the demands of these microorganisms in the market. In this review, are discussing the synergistic application of both cellulose and xylanase enzyme as biocatalyst in various industries. Besides this, the paper also discusses the potential applications and requirements of microbial systems producing such enzymes. The paper highlights the future prospects associated with the further development of these enzymes as a coherent part of various industrial processes.

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Antibodies Targeting Influenza Viruses – A Hope for Universal Vaccine

Succession of physiological stages hallmarks the transcriptomic response of fungus Aspergillus niger to lignocellulose

Gaining insights about the mechanism of lignocellulose degradation by fungi is one of the pathbreaking discoveries for renewable-based biotechnology, specifically for the production of hydrolytic enzymes. Many detailed studies are available which evaluates fungal degradation when the expression levels of CAZyme reach its peak. However, the basic understanding of how fungi survive on lignocelluloses is still under the pipeline. This paper uses Aspergillus niger to explore its responses against six different substrates playing role in biofuel production. The responses to Miscanthus were compared with wheat straw, in isolation and in combination with ionic & hydrothermal feedstock pretreatment.  A metabolic model is mapped using a thorough evaluation of genome-wide transcriptome in combination with defined targeted transcripts and protein analyses. The exposure to different substrates showed enhancement in fatty acid oxidation and lipid metabolism transcripts. Strains with deletion of farA (ortholog of the fatty acid beta-oxidation transcriptional regulator) showed the reduced expression level of genes encoding lignocelluloses degradative CAZyme, indicating the role of beta-oxidation in mediating the adaptation to lignocelluloses. Activation of novel secondary metabolite gene clusters was noticed at the later life stages which are of great consideration due to their importance in synthesis of bioactive compounds. The entire study highlights that substrate composition and structure influence plays a critical role in mediating responses of fungi to lignocellulose. The study also shows the presence of different physiological stages in fungal responses which are mostly conserved across different substrates.

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Cell therapy for central nervous system disorders: Current obstacles to progress

Cell therapy for central nervous system disorders: Current obstacles to progress

No wonder there is an urgent need for new therapeutic developments having clinical application in treating the disorders associated with the central nervous system. In light of this, various studies are under pipeline which focuses on diseases like Parkinson’s, traumatic brain injury, stroke, and many more. Developments have been carried out where cell therapy using induced pluripotent stem cells (iPS) have been used as a breakthrough. The use of induced pluripotent stem cells for neurorestoration through transplantation has paved a new path in the clinical applications treating neurological disorders. Besides this, iPS cells for long have served as a robust biological system for the development of various therapeutic drugs. Thus, providing deeper insights into the pathological conditions and potential mechanism for its treatment. Even after many recent advances in cell therapies using iPS, the mesenchymal stem cells derived from adult tissues remain one of the popular options as the donor for cell transplantation. In this review article, the breakthrough research made using cell therapy for neurological disorders is discussed along with a detailed understanding of the obstacles in clinical application of therapy and subsequent solutions.

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Spray-Dried Hierarchical Aggregates of Iron Oxide Nanoparticles and Their Functionalization for Downstream Processing in Biotechnology.

Spray-Dried Hierarchical Aggregates of Iron Oxide Nanoparticles and Their Functionalization for Downstream Processing in Biotechnology.

Iron oxide nanoparticles are known to be used for their exorbitant properties in a multitude of fields. It has huge applications in mining industries, biomedicine, wastewater treatment, protein purification, food processing and much more. In the present study, researchers tried to unfold the understanding of iron oxide nanoparticles structuring using spray drying method. The study showed the formation of micrometer size aggregates having magnetic property comparable to that of individual nanoparticles. Besides this, researchers found that the superparamagnetic property of the multicore structure formed was not lost. The changes in the aggregates through the addition of silica-based nanoparticles in the suspension showed significant control of the resulting magnetization by altering the iron oxide content. Interestingly, it was observed that the changes in silica content to only 20% brought significant alteration in the morphology of iron oxide nanoparticles. Wherein the inflated like shape of the pure iron oxide nanoparticles was observed to be converted into a spherical structure. These aggregates with different magnetization can be separated in a column under the influence of the magnetic field provided by an attached permanent magnet. These iron oxide aggregates are coupled with a ligand containing nitrilotriacetic acid (NTA) groups that can be used to purify 6-histidine tagged proteins. The present studies show the purification of protein A using this mechanism.  The major benefit of this new purification system under the influence magnetic field is the high degree of recyclability. The system can be used effectively without losing much protein.

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CRISPR/Cas9-mediated gene editing for the development of herbicide-resistant plants.

Application of CRISPR/Cas9-mediated gene editing for the development of herbicide-resistant plants.

Authors: Yun-Jeong Han, Jeong-Il Kim

The development of herbicide-resistant plants is the need of time to help the agriculture economy move ahead. Lack of this trait in plants can impact their growth during herbicide applications leading to loss of crops. Imparting herbicide-resistant traits using genetic engineering will not only help plants/ crops survive better but also control the unwanted weeds. For these reasons, researchers are developing new methods and technologies to pass on herbicide resistance traits in plants. The CRISPR/Cas9 based gene-editing technology is considered to be highly precise and effective to carry out modification at genetic levels offering great efficiency in enhancing the crops. In the present study, the researchers used CRISPR based gene-editing technology to confer herbicide-resistant traits in plants by targeting genes such as 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), acetolactate synthase (ALS), splicing factor 3B subunit 1 (SF3B1), and cellulose synthase A catalytic subunit 3 (CESA3). Besides this, researchers also added additional candidate genes to enhance the herbicide resistance properties of plants through inducing mutations using the CRISPR Cas9 gene-editing tool.

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