Journal of Structural and Functional Genomics
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Differential effects of supplementary affinity tags on the solubility of MBP fusion proteins
Journal of Structural and Functional Genomics - Tập 2 - Trang 83-92 - 2002
It is difficult to imagine any strategy for high-throughput protein expression and purification that does not involve genetically engineered affinity tags. Because of its ability to enhance the solubility and promote the proper folding of its fusion partners, Escherichia coli maltose-binding protein (MBP) is a particularly useful affinity tag. However, not all MBP fusion proteins bind efficiently ......
Computational identification of microRNAs and their targets from the expressed sequence tags of horsegram (Macrotyloma uniflorum (Lam.) Verdc.)
Journal of Structural and Functional Genomics - Tập 11 - Trang 233-240 - 2010
MicroRNAs (miRNAs) are a class of naturally occurring and small non-coding RNA molecules of about 21–25 nucleotides in length. Their main function is to downregulate gene expression in different manners like translational repression, mRNA cleavage and epigenetic modification. To predict new miRNAs in plants different computational approaches have been developed. In the present study, an EST based ......
SUMO fusions and SUMO-specific protease for efficient expression and purification of proteins
Journal of Structural and Functional Genomics - Tập 5 - Trang 75-86 - 2004
SUMO (small ubiquitin-related modifier) modulates protein structure and function by covalently binding to the lysine side chains of the target proteins. Yeast cells contain two SUMO proteases, Ulp1 and Ulp2, that cleave sumoylated proteins in the cell. Ulp1 (SUMO protease 1) processes the SUMO precursor to its mature form and also de-conjugates SUMO from side chain lysines of target proteins. Here......
SOn-column Protein Refolding for Crystallization
Journal of Structural and Functional Genomics - Tập 6 - Trang 177-182 - 2005
One major bottleneck in protein production in Escherichia coli for structural genomics projects is the formation of insoluble protein aggregates (inclusion bodies). The efficient refolding of proteins from inclusion bodies is becoming an important tool that can provide soluble native proteins for structural and functional studies. Here we report an on-column refolding method established at the Ber......
Methods and Results for Semi-automated Cloning Using Integrated Robotics
Journal of Structural and Functional Genomics - Tập 6 - Trang 89-94 - 2005
The Joint Center for Structural Genomics (JCSG) has emphasized automation and parallel processing approaches. Here, we describe automated methods used across the cloning process with results from JCSG projects. The protocols for PCR, restriction digests and ligations, as well as for gel electrophoresis and microtiter plate assays have all been automated. The system has the capacity to routinely pr......
Project management system for structural and functional proteomics: Sesame
Journal of Structural and Functional Genomics - Tập 4 - Trang 11-23 - 2003
A computing infrastructure (Sesame) has been designed to manage and link individual steps in complex projects. Sesame is being developed to support a large-scale structural proteomics pilot project. When complete, the system is expected to manage all steps from target selection to data-bank deposition and report writing. We report here on the design criteria of the Sesame system and on results dem......
Heterologous expression of L. major proteins in S. cerevisiae: a test of solubility, purity, and gene recoding
Journal of Structural and Functional Genomics - Tập 10 - Trang 233-247 - 2009
High level expression of many eukaryotic proteins for structural analysis is likely to require a eukaryotic host since many proteins are either insoluble or lack essential post-translational modifications when expressed in E. coli. The well-studied eukaryote Saccharomyces cerevisiae possesses several attributes of a good expression host: it is simple and inexpensive to culture, has proven genetic ......
Erratum to: The NMR solution structures of the five constituent cold-shock domains (CSD) of the human UNR (upstream of N-ras) protein
Journal of Structural and Functional Genomics - Tập 11 Số 2 - Trang 189-189 - 2010
Solution NMR structures of homeodomains from human proteins ALX4, ZHX1, and CASP8AP2 contribute to the structural coverage of the Human Cancer Protein Interaction Network
Journal of Structural and Functional Genomics - Tập 15 Số 4 - Trang 201-207 - 2014
High-quality solution NMR structures of three homeodomains from human proteins ALX4, ZHX1 and CASP8AP2 were solved. These domains were chosen as targets of a biomedical theme project pursued by the Northeast Structural Genomics Consortium. This project focuses on increasing the structural coverage of human proteins associated with cancer.
Efficient condensed-phase production of perdeuterated soluble and membrane proteins
Journal of Structural and Functional Genomics - Tập 11 Số 2 - Trang 143-154 - 2010
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