Abstract:
:Targeting of secreted and cell-surface proteins to the cell membrane is mediated by a short hydrophobic stretch of amino acids, termed the signal sequence. We have developed a method that detects signal sequences in cDNA fragments based on their ability to redirect a constitutively active mutant of a cytokine receptor to the cell surface, thereby permitting interleukin-3 (IL-3)-independent growth of Ba/F3 cells. Retrovirus-mediated expression of the fusions in IL-3-dependent cells was followed by selection of clones for growth in the absence of IL-3. Infection of cells with 5x10(6) viral particles in a pilot experiment led to the isolation of 150 known and 48 novel cDNA clones, and all the known cDNA clones were found to encode secreted and cell-surface proteins. In addition, we isolated type II membrane proteins, which have not been detected by existing signal sequence trap strategies.
journal_name
Nat Biotechnoljournal_title
Nature biotechnologyauthors
Kojima T,Kitamura Tdoi
10.1038/8666subject
Has Abstractpub_date
1999-05-01 00:00:00pages
487-90issue
5eissn
1087-0156issn
1546-1696journal_volume
17pub_type
杂志文章abstract::We have developed novel nucleic acid probes that recognize and report the presence of specific nucleic acids in homogeneous solutions. These probes undergo a spontaneous fluorogenic conformational change when they hybridize to their targets. Only perfectly complementary targets elicit this response, as hybridization d...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt0396-303
更新日期:1996-03-01 00:00:00
abstract::Of paramount importance for the development of cell therapies to treat diabetes is the production of sufficient numbers of pancreatic endocrine cells that function similarly to primary islets. We have developed a differentiation process that converts human embryonic stem (hES) cells to endocrine cells capable of synth...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt1259
更新日期:2006-11-01 00:00:00
abstract::To enhance milk composition and milk processing efficiency by increasing the casein concentration in milk, we have introduced additional copies of the genes encoding bovine beta- and kappa-casein (CSN2 and CSN3, respectively) into female bovine fibroblasts. Nuclear transfer with four independent donor cell lines resul...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt783
更新日期:2003-02-01 00:00:00
abstract::The site-specific recombinases FLP and Cre are useful for genomic engineering in many living systems. Manipulation of their enzymatic properties offers a means to improve their applicability in different host organisms. We chose to manipulate the thermolability of FLP recombinase. A lacZ-based recombination assay in E...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt0798-657
更新日期:1998-07-01 00:00:00
abstract::We developed a modified flagellar type III secretion apparatus to secrete heterologous polypeptides into the growth medium of Escherichia coli. The secretion was facilitated by fusing the 173-bp untranslated DNA fragment upstream of the gene fliC (encoding flagellin) as well as a transcriptional terminator from fliC, ...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt1077
更新日期:2005-04-01 00:00:00
abstract::In vitro reprogramming of somatic cells into a pluripotent embryonic stem cell-like state has been achieved through retroviral transduction of murine fibroblasts with Oct4, Sox2, c-myc and Klf4. In these experiments, the rare 'induced pluripotent stem' (iPS) cells were isolated by stringent selection for activation of...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt1335
更新日期:2007-10-01 00:00:00
abstract::Systemic lupus erythematosus (SLE) is a multisystem chronic inflammatory disease of unknown etiology that affects many organs, including the kidney. The presence of multiple autoantibodies and other immunological abnormalities point to basic defects in immunoregulatory controls that normally maintain self-tolerance. T...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/77296
更新日期:2000-07-01 00:00:00
abstract::Vaccination with dendritic cells (DCs) pulsed with antigenic peptides derived from various tumor antigens has great, but as yet significantly unrealized, potential in cancer treatment. Here, we describe a strategy for prolonged presentation of an MHC class I-restricted self-peptide on DCs through linkage of it to a ce...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt0202-149
更新日期:2002-02-01 00:00:00
abstract::Functional genomics and proteomics are identifying many potential drug targets for novel therapeutic proteins, and both rational and combinatorial protein engineering methods are available for creating drug candidates. A central challenge is the definition of the most appropriate design criteria, which will benefit cr...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt1064
更新日期:2005-02-01 00:00:00
abstract::The efficiency of both phage display in Escherichia coli and periplasmic expression of recombinant proteins may be limited by the same periplasmic folding steps. To search for E. coli factors that improve the efficiency of both procedures, a library of E. coli proteins was coexpressed in a phagemid vector that contain...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt0498-376
更新日期:1998-04-01 00:00:00
abstract::Circular RNA transcripts were first identified in the early 1990s but knowledge of these species has remained limited, as their study through traditional methods of RNA analysis has been difficult. Now, novel bioinformatic approaches coupled with biochemical enrichment strategies and deep sequencing have allowed compr...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.2890
更新日期:2014-05-01 00:00:00
abstract::Little is known about the potential of plastids to accomplish post-translational modifications of foreign proteins. In the present study we generated transplastomic tobacco plants that accumulate the outer surface lipoprotein A (OspA)-the basic constituent of the first generation monovalent human vaccine against Lyme ...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt1170
更新日期:2006-01-01 00:00:00
abstract::Understanding how proteins and their complex interaction networks convert the genomic information into a dynamic living organism is a fundamental challenge in biological sciences. As an important step towards understanding the systems biology of a complex eukaryote, we cataloged 63% of the predicted Drosophila melanog...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt1300
更新日期:2007-05-01 00:00:00
abstract::High-density short oligonucleotide microarrays have become a widely used tool for measuring gene expression on a large scale. However, details of the mechanism of binding on microarrays remain unclear. Short oligonucleotide probes currently synthesized on microarrays are often ineffective as a result of limited sequen...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt836
更新日期:2003-07-01 00:00:00
abstract::We report a protocol for the rapid identification of DNA-binding proteins. Immobilized DNA probes harboring a specific sequence motif are incubated with cell or nuclear extract. Proteins are analyzed directly off the solid support by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The det...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/12873
更新日期:1999-09-01 00:00:00
abstract::The functional state of a cell is largely determined by the spatiotemporal organization of its proteome. Technologies exist for measuring particular aspects of protein turnover and localization, but comprehensive analysis of protein dynamics across different scales is possible only by combining several methods. Here w...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.2281
更新日期:2012-06-24 00:00:00
abstract::The ability to detect single protein molecules in blood could accelerate the discovery and use of more sensitive diagnostic biomarkers. To detect low-abundance proteins in blood, we captured them on microscopic beads decorated with specific antibodies and then labeled the immunocomplexes (one or zero labeled target pr...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.1641
更新日期:2010-06-01 00:00:00
abstract::Human embryonic stem cells (hESCs) have two properties of interest for the development of cell therapies: self-renewal and the potential to differentiate into all major lineages of somatic cells in the human body. Widespread clinical application of hESC-derived cells will require culture methods that are low-cost, rob...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.1629
更新日期:2010-06-01 00:00:00
abstract::Tissue function depends on hierarchical structures extending from single cells ( approximately 10 microm) to functional subunits (100 microm-1 mm) that coordinate organ functions. Conventional cell culture disperses tissues into single cells while neglecting higher-order processes. The application of semiconductor-dri...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt1361
更新日期:2008-01-01 00:00:00
abstract::One-size-fits-all intravenous delivery of biologics may be giving way to alternate delivery routes that enable safer and more efficient drug administration. Michael Eisenstein reports. ...
journal_title:Nature biotechnology
pub_type: 新闻
doi:10.1038/nbt.1768
更新日期:2011-02-01 00:00:00
abstract::Correct reconstruction of macromolecular structure by cryo-electron microscopy (cryo-EM) relies on accurate determination of the orientation of single-particle images. For small (<100 kDa) DNA-binding proteins, obtaining particle images with sufficiently asymmetric features to correctly guide alignment is challenging....
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/s41587-020-0716-8
更新日期:2020-10-19 00:00:00
abstract::The treatment of asthma and allergic rhinitis using unique, humanized anti-IgE monoclonal antibodies with very particular binding specificities is now supported by the results of multiple phase II and III human clinical studies. The therapeutic efficacy of this approach is attributable to several pharmacological mecha...
journal_title:Nature biotechnology
pub_type: 杂志文章,评审
doi:10.1038/72601
更新日期:2000-02-01 00:00:00
abstract::Isolating antigen-specific T lymphocytes is hampered by the low frequency of the cells and the low affinity between T-cell receptors (TCR) and antigen. We describe the isolation and purification of antigen-specific CD8+ T lymphocytes from mixed T-cell populations. Magnetic beads coated with major histocompatibility co...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt0398-281
更新日期:1998-03-01 00:00:00
abstract::The reactive thiol in cysteine is used for coupling maleimide linkers in the generation of antibody conjugates. To assess the impact of the conjugation site, we engineered cysteines into a therapeutic HER2/neu antibody at three sites differing in solvent accessibility and local charge. The highly solvent-accessible si...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.2108
更新日期:2012-01-22 00:00:00
abstract::We present two algorithms to predict the activity of AsCpf1 guide RNAs. Indel frequencies for 15,000 target sequences were used in a deep-learning framework based on a convolutional neural network to train Seq-deepCpf1. We then incorporated chromatin accessibility information to create the better-performing DeepCpf1 a...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.4061
更新日期:2018-03-01 00:00:00
abstract::DNA charge transport chemistry is found to provide a sensitive method for probing protein-dependent changes in DNA structure and enzymatic reactions. Here we describe the development of an electrochemical assay of protein binding to DNA-modified electrodes based upon the detection of associated perturbations in DNA ba...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt0302-282
更新日期:2002-03-01 00:00:00
abstract::How intrinsic gene-regulatory networks interact with a cell's spatial environment to define its identity remains poorly understood. We developed an approach to distinguish between intrinsic and extrinsic effects on global gene expression by integrating analysis of sequencing-based and imaging-based single-cell transcr...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.4260
更新日期:2018-10-29 00:00:00
abstract::Transplantation of oligodendrocyte precursor cells (OPCs) is a promising potential therapeutic strategy for diseases affecting myelin. However, the derivation of engraftable OPCs from human pluripotent stem cells has proven difficult and primary OPCs are not readily available. Here we report the generation of induced ...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.2564
更新日期:2013-05-01 00:00:00
abstract::Lysosomal storage diseases are treated with human lysosomal enzymes produced in mammalian cells. Such enzyme therapeutics contain relatively low levels of mannose-6-phosphate, which is required to target them to the lysosomes of patient cells. Here we describe a method for increasing mannose-6-phosphate modification o...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.2427
更新日期:2012-12-01 00:00:00
abstract::A human skin allograft injury model in immunodeficient mice, engrafted with human peripheral blood mononuclear cells from a different donor, has been used to test whether reagents that block human T cell CD2 interactions with its principal ligand, LFA-3 (CD58), can inhibit immune reactions in vivo. In this model, huma...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt0897-759
更新日期:1997-08-01 00:00:00