Cloning of S100B cDNA into E. coli under the control of an inducible promoter and its heterologous expression

Abstract

To investigate the structure and functions of the S100B protein in cells and tissues, the heterologous expression of S100B cDNA was studied on
E. coli. A strategy for cloning and expression of S100B cDNA was developed using an efficient and multifunctional online program, Genome Compiler, based on a pBT7-N-His vector system, with a which had a phage T7 RNA polymerase promoter and a cloning and expression site with key elements for control and regulation of transcription and translation. The E. coli strain, BL-21 (DE3) containing the DE3 prophage with the T7 RNA polymerase gene under the control of the lac promoter was used for the expression. The expression of S100B protein was confirmed by the SDS-PAGE procedure, and preliminary studies were performed to optimize the expression procedure. The obtained heterologous expression of the protein with preserved basic physicochemical properties and biological activity opens good prospects for using it as an informative marker of the body state in normal and pathological conditions and as a valuable target of therapy.