Research Topics
Genomes and Genes | James C SamuelsonSummaryAffiliation: New England Biolabs Country: USA Publications
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Publications
Recent developments in difficult protein expression: a guide to E. coli strains, promoters, and relevant host mutationsJames C Samuelson
New England Biolabs, Inc, Ipswich, MA, USA
Methods Mol Biol 705:195-209. 2011..This chapter discusses the properties of many of the E. coli strains available for protein expression in order to facilitate the choice of the best expression host for a particular protein of interest...
Engineering a rare-cutting restriction enzyme: genetic screening and selection of NotI variantsJames C Samuelson
New England Biolabs, Inc 240 County Road, Ipswich, MA 01938, USA
Nucleic Acids Res 34:796-805. 2006..2 x 10(4) U/mg, a 32-fold increase over variant E156K. A comprehensive analysis indicates that the cleavage specificity of M91V/E156K is relaxed to a small set of 8 bp substrates while retaining activity towards the NotI sequence...- The isolation of strand-specific nicking endonucleases from a randomized SapI expression libraryJames C Samuelson
New England Biolabs, Inc, 32 Tozer Road, Beverly, MA 01915, USA
Nucleic Acids Res 32:3661-71. 2004..The nature of the amino acid substitutions found in the selected variants provides evidence that SapI may possess two active sites per monomer. This work presents a framework for establishing the mechanism of SapI DNA cleavage... - Engineering BspQI nicking enzymes and application of N.BspQI in DNA labeling and production of single-strand DNAPenghua Zhang
New England Biolabs, Inc, 240 County Road, Ipswich, MA 01938, USA
Protein Expr Purif 69:226-34. 2010..Restriction mapping and run-off DNA sequencing of digested products from the partially purified enzyme indicated that it is an EarI isoschizomer with 6-bp recognition, which we named CatHI (CTCTTC N1/N4)... - Discovery of natural nicking endonucleases Nb.BsrDI and Nb.BtsI and engineering of top-strand nicking variants from BsrDI and BtsIShuang yong Xu
New England Biolabs, Inc, 240 County Road, Ipswich, MA 01938, USA
Nucleic Acids Res 35:4608-18. 2007..Top-strand specific nicking variants, Nt.BsrDI and Nt.BtsI, were successfully engineered by combining the catalytic-deficient B subunit with wild-type A subunit... 
Directed evolution of restriction endonuclease BstYI to achieve increased substrate specificityJames C Samuelson
New England Biolabs, 32 Tozer Road, Beverly, MA 01915, USA
J Mol Biol 319:673-83. 2002..Moreover, cleavage of the GGATCC sequence is no longer detected. This study provides further evidence that laboratory evolution strategies offer a powerful alternative to structure-guided protein design...- Engineering Escherichia coli BL21(DE3) derivative strains to minimize E. coli protein contamination after purification by immobilized metal affinity chromatographyCarine Robichon
New England Biolabs, Inc, Gene Expression Division, 240 County Road, Ipswich, MA 01938, USA
Appl Environ Microbiol 77:4634-46. 2011..The "NiCo" strains uniquely complement existing methods for improving the purity of recombinant His-tagged protein... - Engineering strand-specific DNA nicking enzymes from the type IIS restriction endonucleases BsaI, BsmBI, and BsmAIZhenyu Zhu
New England Biolabs, Inc. 32 Tozer Road, Beverly, MA 01915, USA
J Mol Biol 337:573-83. 2004..This work provides strong evidence that some type IIS restriction endonucleases carry two separate active sites. When one of the active sites is inactivated, the type IIS restriction endonuclease may nick only one strand... - Rational design of a fusion partner for membrane protein expression in E. coliJianying Luo
New England Biolabs, Inc Gene Expression Division, Ipswich, Massachusetts 01938, USA
Protein Sci 18:1735-44. 2009..We anticipate that this optimized expression system will aid in the isolation and study of various recombinant forms of membrane-associated protein... - BstYI bound to noncognate DNA reveals a "hemispecific" complex: implications for DNA scanningSharon A Townson
Department of Structural and Chemical Biology, Mount Sinai School of Medicine, 1425 Madison Avenue, New York, NY 10029, USA
Structure 15:449-59. 2007..Taken together, the structure provides a snapshot of an enzyme in a "paused" intermediate state that may be part of a more general mechanism of scanning DNA... - Implications for switching restriction enzyme specificities from the structure of BstYI bound to a BglII DNA sequenceSharon A Townson
Structural Biology Program, Department of Physiology and Biophysics, Mount Sinai School of Medicine, 1425 Madison Avenue, New York, New York 10029, USA
Structure 13:791-801. 2005..Taken together, the structure reveals a mechanism of degenerate DNA recognition and offers insights into the possibilities and limitations in altering specificities of closely related restriction enzymes... - Crystal structure of BstYI at 1.85A resolution: a thermophilic restriction endonuclease with overlapping specificities to BamHI and BglIISharon A Townson
Structural Biology Program, Department of Physiology and Biophysics, Mount Sinai School of Medicine, 1425 Madison Avenue, New York, NY 10029, USA
J Mol Biol 338:725-33. 2004..This arm substructure may underlie the thermostability of BstYI. We identify putative DNA recognition residues and speculate as to how this enzyme achieves a "relaxed" DNA specificity... - Direct interaction of YidC with the Sec-independent Pf3 coat protein during its membrane protein insertionMinyong Chen
Department of Chemistry, Ohio State Biochemistry Program, and Protein Research Group, The Ohio State University, Columbus, Ohio 43210, USA
J Biol Chem 277:7670-5. 2002..Our study demonstrates that YidC can directly interact with a Sec-independent membrane protein, and the role of YidC is at the stage of folding the Pf3 protein into a transmembrane configuration...