Expression of tomato thymidine kinase 1 by means of the baculovirus expression vector system

ABSTRACT

Tomato thymidine kinase 1 (ToTK1) is a deoxyribonucleoside kinase (dNK) that has been subject to study because of its potential to phosphorylate the nucleoside analogue 3-azido-2,3-dideoxythymidine (azidothymidine, AZT) equally well as its natural substrate thymidine (dThd). The combination of ToTK1 and AZT has been tested in two animal studies for its efficiency and use in suicide gene therapy for malignant glioma. The determination of the 3D structure of ToTK1 might shed light on the structure–function relationships of nucleoside activation by this enzyme and thereby show routes toward further improvement of ToTK1 and other TK1-like dNKs for suicide gene therapy. Here we report the successful expression of both full-length ToTK1 and a C-terminal truncated ToTK1 in Spodoptera frugiperda and Trichoplusia ni insect cells using the baculovirus expression vector system. This constitutes a further step on the road to determine the 3D structure of the first TK1 of plant origin, but also an enzyme with great potential for dNK-mediated suicide gene therapy.     

NSort/DB:An Intranuclear Compartment Protein Database

Distinct substructures within the nucleus are associated with a wide variety of important nuclear processes.Structures such as chromatin and nuclear pores have specific roles,while others such as Cajal bodies are more functionally varied.Understanding the roles of these membraneless intra-nuclear compartments requires extensive data sets covering nuclear and compartment-associated proteins.NSort/DB is a database providing access to intra-or sub-nuclear compartment associations for the mouse nuclear proteome.Based on resources ranging from large-scale curated data sets to detailed experiments,this data set provides a high-quality set of annotations of non-exclusive association of nuclear proteins with structures such as promyelocytic leukaemia bodies and chromatin.The database is searchable by protein identifier or compartment,and has a documented web service API.The search interface,web service and data download are all freely available online at http://www.nsort.org/db/.Availability of this data set will enable systematic analyses of the protein complements of nuclear compartments,improving our understanding of the diverse functional repertoire of these structures.     

Existence of sodium bicarbonate enhanced bioelectricity generation on Chlorella sp. biofilm in a Biophotovoltaic (BPV) system

Journal of Applied Phycology - Sodium bicarbonate (NaHCO3) is a low-cost inorganic carbon source, highly soluble in water and widely used in microalgae research. Using carbon dioxide (CO2) gas in...     

Carbon quantum dots fluorescence quenching for potassium optode construction

Photophysical phenomena associated with carbon nanoparticles in combination with biocompatibility and readily functionalizable properties have attracted significant interest for sensing and imaging applications. A potassium ion optode based on the fluorescence quenching of carbon quantum dots (CQDs) was constructed. The CQDs were synthesized using a microwave method, citric acid and 2,2′‐(ethylene‐dioxy)bis(ethylamine). A quantum yield of 7.1% was calculated for the synthesized CQDs. A linear dynamic range of about one‐order of magnitude with a correlation coefficient of 0.99 was obtained. The optode was applied on real samples and a 0.60–1.60% error range was obtained relative to the ion‐selective electrode.