Analytische Elektronenmikroskopie in Molekularbiologie und Biotechnologie Abschlussbericht

Abstract

A lot of molecular biological analysis centre on protein DNA interactions being essential for gene activation. An innovation step in the field of Pol I-Promoter associated transscription factors made it for the first time possible to examine the structures of such interactions. The intention was to map DNA in small DNA-protein-complexes via phosphate detection by means of high-resolution electron microscopy (EM), especially the electron energy loss spectroscopy, and to optimize the image possibilities of molecular biologically important marking elements - B, N, F, Fe, J -. A novel instrument configuration with highly sensitive detection on basis of a SSCCD-camera with image analysis and the LEO 912 OMEGA with integrated energy dispersive filter should have been developed for this purpose. Easy handling for molecular biological applications had been an important aim of this development. The methods for P-detection having been described so far do not allow reliable results. More intensive basic analysis had therefore been necessary. The DNA imaging had not fully been reached. However, a two-window-process had been developed and applied for a patent determining the background to the phosphor element signal via a mass thickness standard. For comparison of various methods for evaluation of the P-signal a method had been prepared covering and evaluating systematic and statistical errors of the measurement using RNA in Turnip-Yellow-Mosaic viruses as example. This optimized instrument system is a good tool for getting to know the structural correlative functions of differential gene activity in a better way. (orig.)SIGLEAvailable from TIB Hannover: F99B314 / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekBundesministerium fuer Bildung, Wissenschaft, Forschung und Technologie, Bonn (Germany)DEGerman