Dihydrostreptomycin [HRP]

Glycine-rich RNA-binding proteins (GR-RBPs) have been explored in different plant species for their role in development and environmental stress responses. NtGR-RBP1, a tobacco glycine-rich RNA-binding protein has recently been suggested as RNA chaperone with reference to protective role in bacteria upon exposure to various stresses. In the present study, we aimed to test the roles of NtGR-RBP1 in plants. The cDNA sequence of NtGR-RBP1 was amplified and cloned into entry vector using pENTR/D-TOPO cloning kit followed by sub-cloning into a plant GATEWAY (R) expression vector resulting in construction of pH7WG2-NtGR-RBP1. NtGR-RBP1 recombinants were confirmed through colony PCR and sequencing. Further, pH7WG2-NtGR-RBP1 was cloned into Agrobacterium tumefaciens AGL1 competent cells and confirmed by PCR after screening on streptomycin and hygromycin. The culture was subsequently used for the transformation of Nicotiana tabacum cv samsun by Agrobacterium-mediated plant transformation using the leaf disc method. Explants were screened using hygromycin as selection marker and later confirmed through PCR. Seed germination assay showed that NtGR-RBP1 had significant role in response to cold, heat and drought stress, while salt stress has negative effect on germination. Transformed plants will be used further for in depth functional characterization of NtGR-RBP1.

A low cost, sensitive and selective electrochemical imprinted biosensor for horseradish peroxidase-conjugated prostate-specific antigen (PSA) was designed and prepared based on the combination of Toluidine blue (TB) and self-assembly surface molecular imprinting technique. Poly toluidine blue [P(TB)] provided high surface area for dense loading of HRP-PSA antibody on GCE surface. P(TB), as supporting material, could effectively enhance imprinting efficiency and the electrode conductivity and facilitate electron transfer. The imprinted biosensor was characterized through Field emission scanning electron microscopy (FE-SEM), Energy-dispersive X-ray spectroscopy (EDX) and electrochemical methods. The proposed biosensor indicates very highly electrocatalytical activity for the reduction of hydrogen peroxide (H2O2). Also, engineered biosensor was used for determination of H2O2 by different electrochemical techniques including differential pulse voltammetry, square wave voltammetry and chronoamperometry. Under the optimized conditions, the proposed bio-imprinted polymer exhibit excellent electrocatalytical activity toward the reduction of H2O2 with wide linear range of 0.001 to 40 mM and a low limit of quantification (LLOQ) of 1 mu M.