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551-0379-00L 6 Credits BSC D-BIOL

Exploring Chromatin Structure and Biophysics

Lecturers & Examiners: Prof. em. Dr. Timothy J. Richmond
VVZ CR n/a

Last Updated: 2026-02-05 15:23:45

Abstract

Various methodologies, from protein expression to X-ray analysis, are applied to structural and biophysical studies of chromatin.

Objective

Potential topics of study: 1) Understand why chromatin structural and biophysical studies are important, and specifically why the experiments to be performed are of interest. Learn how to achieve the technical requirements for structural studies on macromolecular complexes (milligram quantities; high concentration, purity and stability; limited conformational homogeneity). Learn what questions the available techniques can answer. 2) Procedures for protein expression (cloning, PCR, in vitro recombination, cell growth) and purification (chromatography). 3) Procedures for in vitro assembly of macromolecular complexes based on the nucleosome core particle or a dodecanucleosome. 4) Biophysical characterization of the nucleosome core particle or dodecanucleosome (native gels, analytical ultracentrifugation). 5) Crystallization techniques. 6) Preliminary X-ray analysis.

Content

Potential types of experiments: I. Design histone mutants that effect various binding interfaces (DNA, remodeling and modification factors). Use QuickChange to produce an expression vector containing a point mutant. Use the Infusion recombination system for cloning and mutation. II. Express and purify mutant histone proteins. Assemble them into nucleosome core particles and dodecanucleosomes. Alternatively, assemble a 147 bp DNA fragment of an sequence of interest into nucleosome core particles. III. Analyze wild-type and mutant nucleosome core particles and dodecanucleosomes by using native-gel mobility assays. Use Sca I digest, native gel mobility assay and sedimentation velocity for analysis of dodecanucleosomes. IV. Crystallization of the nucleosome core particle, followed by X-ray data analysis using lab equipment. With incorporation of cysteine, the mutant could be visualized using methylmercury.

General Information

Language
English
Levels
BSC
Frequency
Yearly recurring

Examination

Type
graded semester performance

Course Components

Type Title Time & Place Hours
lecture with exercise Exploring Chromatin Structure and Biophysics
Permission from lecturers required for all students. Block course held in each quarter of the semester. (restricted to 2 candidates per quarter / 8 candidates per semester and year) Blockkurs im 1. Viertel beginnt bereits am 16.09.2008
  • Tue 12:45-16:30 (HPK F 3)
  • Wed 07:45-16:30 (HPK F 3)
  • Thu 07:45-16:30 (HPK F 3)
  • Fri 07:45-16:30 (HPK F 3)
  • 16.09 Date 12:45-16:30 (HPK F 3)
100 h semesterly

Offered In