Cloning and expression of human neuron-specific enolase cDNA in Escherichia coli.

cDNA fragment encoding neuron-specific enolase was amplified from the cDNA library of human brain. Then the fragment was cloned for expression in E. coli using the vector pET28-a.

Immunofluorescent analysis of connexin-43 using monoclonal antibodies to its extracellular domain.

Immunofluorescent analysis of connexin-43 was carried out on preparations of fixed and living cultures of rat and human glioma cells, HEK293 cells, and frozen sections of the rat brain with experimental glioma using monoclonal antibodies to recombinant extracellular fragment of connexin-43 (E2 second extracellular loop). These monoclonal antibodies visualized membrane and cytoplasmic pools of connexin-43 in preparations fixed with paraformaldehyde. Incubation of monoclonal antibodies to E2 extracellular loop with living cells led to visualization of only connexin hemichannels on cell membranes.

Immunoliposomal containers as systems of directed transport of minor interfering RNA into Schwann cells.

Immunoliposomal container system for targeted delivery of minor interfering RNA into Schwann cells was developed. Monoclonal antibodies to myelin basic protein served as the vector. Analysis by sandwich ELISA of myelin basic protein showed specific suppression of the target protein gene expression in Schwann cells after incubation with PEG-conjugated immunoliposomes loaded with minor interfering RNA blocking the synthesis of myelin basic protein.

Cloning and expression of rat GFAP cDNA in Escherichia coli.

A cDNA fragment encoding GFAP was amplified by reverse transcription PCR from total mRNA isolated from primary culture of rat astrocytes and cloned for expression in Escherichia coli using pET-28a vector. High level of GFAP expression was confirmed by SDS-PAGE, while immunochemical identity was verified by immunoblotting. The constructed producer strain is a cheap source of GFAP and can be used for diagnostic purposes.