Molecular mechanism of biophylaxis for adverse factor in working environment

It has been known that there exist some specific proteins in the pathway by which adverse environmental components are transferred to genes mediating the biophylaxis mechanism. Therefore, it seemed that information exchange might be made through interaction between such proteins and unknown proteins, and this step would be the key step of biophylaxis. In this study, construction of a recombinant protein was attempted to develop a detection method for such protein-protein interaction using radioisotopes. First, vectors that can express ZRF protein with S-tag at the N-terminal end in E.coli were constructed (pZRF29, pZRF30) and E.coli transfected with the vector was cultured to collect the recombinant protein. Thus obtained recombinant ZRF protein was analyzed by polyacrylamide gel electrophoresis and detection was made by Coomassie staining. Using the two vectors, ZRF protein was produced. The protein could be obtained at much higher efficiency in the present system than the previous system. However, the expression level of β-galactosidase as the control was still higher than that of the ZRF protein. This would be due to the higher stability of β- galactosidase, suggesting that further improvement might be obtainable by stabilizing the ZRF protein at the time of expression. It was thought necessary to choose the optimum conditions of the system in respects of cell culture and induction of the recombinant protein as well as protein extraction. (M.N.)