To begin to address the functional interactions between constitutively released nucleotides, ectonucleotidase activity, and P2Y receptor-promoted signaling responses, we engineered the human P2Y(1) receptor in a fusion protein with a member of the ectonucleoside triphosphate diphosphohydrolase family, NTPDase1. Membranes prepared from Chinese hamster ovary (CHO)-K1 cells stably expressing eithe...

conclusions the gp40/15 gene, which was cloned in the pet28a+, was successfully expressed and produced in e. coli. therefore, this protein can be used in future studies to develop recombinant vaccines and diagnostic kits. methods in this experimental study, the gp40/15 gene sequence was extracted from genbank (no. af155624) and cloned in the pet28a+ plasmid. colony polymerase chain reaction (pc...

this paper reviews recent strategies used for increasing specific yield and productivity in high cell density cultures. high cell density cultures offer an efficient means for the economical production of recombinant proteins. however, there are still some challenges associated with high cell density cultivation (hcdc) techniques. a variety of strategies in several aspects including host design...

Herpes simplex virus 1 (HSV-1) unspliced 8.3 latency associated transcript (LAT), which located in the long repeat sequences, has been shown to contain at least 16 open reading frames (ORF: A-P). One of these ORF, ORF P, maps almost entirely antisense to HSV-1 neurovirulence gene, ICP34.5. Both ORF P and ICP34.5 are located in the long repeat and are antisense overlapping genes. Therefore, in O...

Chimeric fluorescent fusion proteins have been employed as a powerful tool to reveal the subcellular localizations and dynamics of proteins in living cells. Co-expression of a fluorescent fusion protein with well-known organelle markers in the same cell is especially useful in revealing its spatial and temporal functions of the protein in question. However, the conventional methods for co-expre...

With the plasmid DNA from a clinical isolate of enterotoxigenic Escherichia coli (ETEC) H 10407 as template, PCR-mediated cloning of the sequence encoding the heat-labile toxin B subunit (L T -B) has been carried out. Then this sequence was recloned into the pTrc 99A and pET23a expression vectors to give the pJasmids pTRCLTB and pETLTB, respectively. After induction, the former plasmid provides...

BACKGROUND: Antibody fragments are molecules widely used for diagnosis and therapy. A large amount of protein is frequently required for such applications. New approaches using folding reporter enzymes have recently been proposed to increase soluble expression of foreign proteins in Escherichia coli. To date, these methods have only been used to screen for proteins with better folding propertie...

AIM To express the fusion protein of enhanced green fluorescent protein (EGFP) with the light chain variable domain of the neutralizing monoclonal antibody MA18/7 (mAb) against hepatitis B virus in E.coli, and determine its bioactivity. METHODS The EGFP gene was cloned into vector pTO-T7 to construct an expression vector. And then according to ORF gene, MA18/7-V(L) was inserted into the 5' te...

prohibitins are chaperone proteins highly conserved among eukaryotes. many different functions have been attributed to prohibitins depending on their tissue expression and sub-cellular localization. prohibitins play a major role in mitochondrial physiology, regulating the stability and the processing of both respiratory chain complex subunits and opa1, a mitochondrial fusion regulating protein....

background: bacterial superantigen staphylococcal enterotoxins (ses), has stimulated polyclonal t cells irrespective of their antigen specificity, resulted a massive release of cytokines, and suggested that they could be assigned as a candidate of new antitumor agents. recent attempts have done to specifically target superantigens towards tumors, subsequently monoclonal antibodies and tumor-rel...