Biologie moléculaire : accès libre

Several biological functions and eukaryotic proteomes are interrelated and they depend upon protein function and analysis.Protein function and interaction is a biological process.. Deviations in these interactions can occur. In order to overcome this problem, scientists have devised a fusion platform called HaloTag technology. It is a technique used for functional analysis of proteins. This technique involves development of a 33kDa HaloTag that is fused to protein of interest and development of HaloTag ligand.When the HaloTag fusion protein tag and ligand come in contact, a covalent bond is formed rapidly. The covalent bond formed is irreversible so it ensures efficient isolation and purification of target protein. Variety of ligands is available for analysis of protein.Proteins always react differently to the new therapeutic products and it is unpredictable to discover their role after interacting with the ligand. HaloTag technology is used for analysis of protein-protein interactions, protein DNA interactions, protein functional analysis, enzyme immobilization and tumor cell detection. HaloTag technology evaluated the function in a way that it captures only protein of interest that is captured and purified.

Quorum sensing is a cell-to-cell signaling process that allows bacteria to collectively control gene expression, thereby harmonizing activities that are productive only at a high population density such as biofilm formation and virulence factors production. This process is accomplished through the production, secretion, and detection of small chemical signals called autoinducers. Autoinducer-2 kinase (LsrK) is the kinase protein present in the quorum sensing system. Generally; kinase protein has been implicated in the progression of a wide range of diseases, making these signaling molecules key therapeutic targets for pharmaceutical development. Hence, we engrossed on LsrK; Gene was cloned into pET30b (+) with specific primers and over expressed under control of the T7 promoter using E. coli BL21 (DE3) bacterial system with 0.1mM IPTG with LB media at 37°C. Denatured LsrK was obtained after treatment GnHCl. These efficient and optimized methods were supportive for refolding and crystallization of S. typhi LsrK.