Suitable vectors for HTP cloning are described and results from their use in _expression_ screening and protein-production pipelines are reported. Of these, yeast and baculovirus-infected insect cells currently represent the _expression_ systems of choice for structural biologists. Some individual projects and developments are discussed in detail, while those that are covered elsewhere in this issue are treated more briefly.
The major obstacle turned out to be poor _expression_ and low solubility. The first is the quality assessment of the produced protein preparations.
However, prokaryotic based _expression_ systems fail to generate correctly folded functional forms of many proteins and hence a variety of eukaryotic based _expression_ systems have been developed. Although important biologically, for the structural biologist disordered regions of proteins can be disastrous even preventing successful structure determination.
Target analysis by biophysical clustering based on pI and hydropathy has provided useful information for future target-selection strategies.
Papers on the crystallization of biological molecules will be accepted providing that these focus on new methods or other features that are of general importance or applicability.
Papers on crystallographic methods should be oriented towards biological crystallography, and may include new approaches to any aspect of structure determination or analysis. Of these, yeast and baculovirus-infected insect cells currently represent the _expression_ systems of choice for structural biologists.
Some individual projects and developments are discussed in detail, while those that are covered elsewhere in this issue are treated more briefly.
However, prokaryotic based _expression_ systems fail to generate correctly folded functional forms of many proteins and hence a variety of eukaryotic based _expression_ systems have been developed. However, prokaryotic based _expression_ systems fail to generate correctly folded functional forms of many proteins and hence a variety of eukaryotic based _expression_ systems have been developed.
SPINE was able to bring together developers of software with users from the partner laboratories.
The technological developments and lessons learned from this project are discussed.
Development areas were broken down into workpackages and this article gives an overview of ongoing activity in the bioinformatics workpackage. Hence, it is essential to perform an extensive quality assessment of the protein preparations prior to crystallization and to use the results in the evaluation of the process.
Here, protocols for a simple, fast and affordable method for transient protein _expression_ in mammalian cells are reported.
coli and a database system dedicated to handle co-_expression_ data are described. Although important biologically, for the structural biologist disordered regions of proteins can be disastrous even preventing successful structure determination. Six human proteins were used to optimize the GRETA fermentation processes and to compare these processes with typical baffled-flask protocols used in structural genomics projects. SPINE was able to bring together developers of software with users from the partner laboratories.
anthracis, established and developed HTP cloning and _expression_ technologies and tested HTP pipelines.
Target analysis by biophysical clustering based on pI and hydropathy has provided useful information for future target-selection strategies.