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 Escherichia coli was used for selection in a protein evolution strategy that relied on error-prone PCR and DNA shuffling. Improved FLP recombinases were identified through cycles of increasing stringency imposed by both raising temperature and reducing protein expression, combined with repetitive cycles of screening at the same stringency to enrich for clones with improved fitness. An eighth generation clone (termed FLPe) showed improved properties in E. coli, in vitro, in human 293- and mouse ES-cells.
journal_name
Nat Biotechnoljournal_title
Nature biotechnologyauthors
Buchholz F,Angrand PO,Stewart AFdoi
10.1038/nbt0798-657subject
Has Abstractpub_date
1998-07-01 00:00:00pages
657-62issue
7eissn
1087-0156issn
1546-1696journal_volume
16pub_type
杂志文章abstract::A bacteriophage lambda surface expression system, lambda foo, was used for epitope mapping of human galectin-3. We constructed random epitope and peptide libraries and compared their efficiencies in the mapping. The galectin-3 cDNA was randomly digested by DNase 1 to make random epitope libraries. The libraries were s...
journal_title:Nature biotechnology
pub_type: 杂志文章
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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
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abstract::As the cost of human genome sequencing plunges and large-scale genome-phenotype studies become possible, society should do more to reward those individuals who choose to disclose their data, despite the risks. ...
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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. ...
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abstract::The biological and physical sciences share a common interest in small structures (the definition of 'small' depends on the application, but can range from 1 nm to 1 mm). A vigorous trade across the borders of these areas of science is developing around new materials and tools (largely from the physical sciences) and n...
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abstract::Multiple sequence alignments (MSAs) are used for structural1,2 and evolutionary predictions1,2, but the complexity of aligning large datasets requires the use of approximate solutions3, including the progressive algorithm4. Progressive MSA methods start by aligning the most similar sequences and subsequently incorpora...
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abstract::An amendment to this paper has been published and can be accessed via a link at the top of the paper. ...
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journal_title:Nature biotechnology
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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...
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doi:10.1038/82414
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abstract::The epicardium and its derivatives provide trophic and structural support for the developing and adult heart. Here we tested the ability of human embryonic stem cell (hESC)-derived epicardium to augment the structure and function of engineered heart tissue in vitro and to improve efficacy of hESC-cardiomyocyte grafts ...
journal_title:Nature biotechnology
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pub_type: 杂志文章,评审
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更新日期:2002-06-01 00:00:00
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journal_title:Nature biotechnology
pub_type:
doi:10.1038/72013
更新日期:2000-01-01 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::The ability to produce monoclonal antibodies (Mabs) in plants offers the opportunity for the development of an inexpensive method of mucosal immunoprotection against sexually transmitted diseases. To investigate the suitability of plant-expressed Mabs for vaginal preventive applications, we compared a humanized anti-h...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/4344
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abstract::Escherichia coli-based artificial chromosomes have become important tools for physical mapping and sequencing in various genome projects. The lack of a general method to modify these large bacterial clones, however, has limited their utility in functional studies. We developed a simple method to modify bacterial artif...
journal_title:Nature biotechnology
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doi:10.1038/nbt0997-859
更新日期:1997-09-01 00:00:00
abstract::Despite advances in DNA sequencing technology, assembly of complex genomes remains a major challenge, particularly for genomes sequenced using short reads, which yield highly fragmented assemblies. Here we show that genome-wide in vivo chromatin interaction frequency data, which are measurable with chromosome conforma...
journal_title:Nature biotechnology
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doi:10.1038/nbt.2768
更新日期:2013-12-01 00:00:00
abstract::Evolution of eukaryotes is mediated by sexual recombination of parental genomes. Crossovers occur in random, but homologous, positions at a frequency that depends on DNA length. As exons occupy only 1% of the human genome and introns about 24%, by far most of the crossovers occur between exons, rather than inside. The...
journal_title:Nature biotechnology
pub_type: 杂志文章,评审
doi:10.1038/88084
更新日期:2001-05-01 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
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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...
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更新日期:2021-01-01 00:00:00
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journal_title:Nature biotechnology
pub_type: 杂志文章
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更新日期:1996-08-01 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
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doi:10.1038/nbt1077
更新日期:2005-04-01 00:00:00
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journal_title:Nature biotechnology
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更新日期:2011-10-09 00:00:00
abstract::L-Ascorbic acid (vitamin C) in fruits and vegetables is an essential component of human nutrition. Surprisingly, only limited information is available about the pathway(s) leading to its biosynthesis in plants. Here, we report the isolation and characterization of GalUR, a gene from strawberry that encodes an NADPH-de...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt777
更新日期:2003-02-01 00:00:00
abstract::Transplantation of pancreatic progenitors or insulin-secreting cells derived from human embryonic stem cells (hESCs) has been proposed as a therapy for diabetes. We describe a seven-stage protocol that efficiently converts hESCs into insulin-producing cells. Stage (S) 7 cells expressed key markers of mature pancreatic...
journal_title:Nature biotechnology
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doi:10.1038/nbt.3033
更新日期:2014-11-01 00:00:00