A study on cloning and expression of HIV-1 NEF protein in HEK 293 cells by transient transfection

HIV continues to be a major global public health issue, there were approximately 34 million people living with HIV in 2011. However, the development of anti-viral has blunted the AIDS epidemic in the Western world but globally the epidemic has not been curtailed. Nef is one of these accessory genes that are only present within HIV and SIV genome and thought to play a key role in the progression to AIDS. The given its central role in HIV pathogenesis, Nef considered as a potential anti-viral target for preventing or at least delaying pathogenesis. The biologically active 27 kDa myristoylated Nef protein expressed from HEK 293 cells is a protein model to be used for significantly specific antibody production to lower the pathogenicity of HIV infection. To express the this protein model, pQBI-6His a mammalian expression vector constructed with base pair 5504 to express 627 bp HIV-1 Nef under CMV promoter. It shows that targeted 27 kDa HIV-1 Nef was not successfully expressed in HEK 293 cells either in transient transfection when transfected. However, nontargeted HIV-1 Nef was detected in western blot by anti-Nef (anti mouse) manufactured by Thermo scientific. The ability of not expressing the targeted myristoylated 27 kDa nef protein was to various unpreventable factors due to time limitation and lacking of skills in the filed cloning and cell culture

Ultra-sensitive biosensor based on genetically engineered acetylcholinesterase immobilized in poly (vinyl alcohol)/Fe–Ni alloy nanocomposite for phosmet detection in olive oil

International audienceAn ultra-sensitive screen-printed biosensor was successfully developed for phosmet detection in olive oil, based on a genetically-engineered acetylcholinesterase (AChE) immobilized in a azide-unit water-pendant polyvinyl alcohol (PVA-AWP)/Fe-Ni alloy nanocomposite. Fe-Ni not only allowed amplifying the response current but also lowering the applied potential from 80 mV to 30 mV vs Ag/AgCl. The biosensor showed a very good analytical performance for phosmet detection, with a detection limit of 0.1 nM. This detection limit is lower than the allowable concentrations set by international regulations. In addition to the good reproducibility, operational and storage stability, the developed biosensor was successfully used for the determination of phosmet in olive oil samples without any laborious pre-treatment. The phosmet recovery rate was about 96% after a simple liquid-liquid extraction

Repurposing of sitagliptin-melittin optimized nanoformula against sars-cov-2: Antiviral screening and molecular docking studies

Abstract: The outbreak of the COVID-19 pandemic in China has become an urgent health and eco-nomic challenge. The objective of the current work was to evaluate the efficacy of the combined complex of Sitagliptin (SIT) with melittin (MEL) against SARS-CoV-2 virus. SIT-MEL nano-conju-gates were optimized by a full three-factor bi-level (23) factorial design. In addition, SIT concentration (mM, X1), MEL concentration (mM, X2), and pH (X3) were selected as the critical factors. Particle size (nm, Y1) and zeta potential (mV, Y2) were assessed as responses. Characterization of the optimized formula for Fourier-transformed infrared (FTIR) was carried out. The optimized formula showed particle size and zeta potential values of 77.42 nm and 27.67 mV, respectively. When compared with SIT and MEL, the combination of SIT-MEL complex has shown anti-viral potential against isolate of SARS-CoV-2 with IC50 values of 8.439 μM with significant improvement (p < 0.001). In addition, the complex showed IC50 in vitro 3CL-protease inhibition with IC50 7.216 µM. Molecular docking has revealed that formula components have good predicted pocket accommodation of the SARS-CoV-2 3-CL protease. An optimized formulation of SIT-MEL could guarantee both enhanced delivery to the target cells and the enhanced cellular uptake with promising activities against SARS-CoV-2. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]