Abstract:
:Phage display technology involves the display of proteins or peptides, as coat protein fusions, on the surface of a phage or phagemid particles. Using standard technology, helper phage are essential for the replication and assembly of phagemid particles, during library production and biopanning. We have eliminated the need to add helper phage by using 'bacterial packaging cell lines' that provide the same functions. These cell lines contain M13-based helper plasmids that express phage packaging proteins which assemble phagemid particles as efficiently as helper phage, but without helper phage contamination. This results in genetically pure phagemid particle preparations. Furthermore, by using constructs differing in the form of gene 3 that they contain, we have shown that the display, from a single library, can be modulated between monovalent (phagemid-like) and multivalent display (phage-like) without any further engineering. These packaging cells eliminate the use of helper phage from phagemid-based selection protocols; reducing the amount of technical preparation, facilitating automation, optimizing selections by matching display levels to diversity, and effectively using the packaged phagemid particles as means to transfer genetic information at an efficiency approaching 100%.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Chasteen L,Ayriss J,Pavlik P,Bradbury ARdoi
10.1093/nar/gkl772subject
Has Abstractpub_date
2006-01-01 00:00:00pages
e145issue
21eissn
0305-1048issn
1362-4962pii
gkl772journal_volume
34pub_type
杂志文章abstract::We present a theory of pluralistic and stochastic gene regulation. To bridge the gap between empirical studies and mathematical models, we integrate pre-existing observations with our meta-analyses of the ENCODE ChIP-Seq experiments. Earlier evidence includes fluctuations in levels, location, activity, and binding of ...
journal_title:Nucleic acids research
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abstract::A new method for specific reamplification of DDRT-PCR products is presented. After transient ligation of the primary DDRT-PCR fragments into a T-vector, the cDNAs of interest were reamplified by hemi-nested PCR and thermally asymmetric cycles. In contrast to the originally described protocol, this method of reamplific...
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更新日期:2020-04-17 00:00:00
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
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更新日期:2007-01-01 00:00:00
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journal_title:Nucleic acids research
pub_type: 杂志文章
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journal_title:Nucleic acids research
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abstract::The crystal structure of d(CATGGGCCCATG)(2) shows unique stacking patterns of a stable B<-->A-DNA intermediate. We evaluated intrinsic base stacking energies in this crystal structure using an ab initio quantum mechanical method. We found that all crystal base pair steps have stacking energies close to their values in...
journal_title:Nucleic acids research
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doi:10.1093/nar/28.24.4893
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journal_title:Nucleic acids research
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更新日期:2009-04-01 00:00:00
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更新日期:1987-08-11 00:00:00
abstract::A screening for human proteins capable of interacting with influenza virus NS1 has been carried out using the two-hybrid genetic trap in yeast. A cDNA corresponding to the human homologue of Drosophila melanogaster Staufen protein (hStaufen) was isolated that fulfilled all genetic controls of the two-hybrid protocol. ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/27.11.2241
更新日期:1999-06-01 00:00:00
abstract::Competition binding and UV melting studies of a DNA model system consisting of three, four or five mutually complementary oligonucleotides demonstrate that unpaired bases at the branch point stabilize three- and five-way junction loops but destabilize four-way junctions. The inclusion of unpaired nucleotides permits t...
journal_title:Nucleic acids research
pub_type: 杂志文章
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journal_title:Nucleic acids research
pub_type: 评论,杂志文章
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更新日期:2003-09-15 00:00:00
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更新日期:2015-10-30 00:00:00
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journal_title:Nucleic acids research
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更新日期:2008-10-01 00:00:00
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journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:2014-01-01 00:00:00
abstract::Human stem cells are promising sources for regenerative therapy. To ensure safety of future therapeutic applications, the differentiation potency of stem cells has to be tested and be widely opened to the public. The potency is generally assessed by teratoma formation comprising differentiated cells from all three ger...
journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:2016-01-04 00:00:00
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更新日期:2020-08-20 00:00:00
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journal_title:Nucleic acids research
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更新日期:1977-11-01 00:00:00
abstract::50 to 55% of tobacco and barley nuclear DNA is accessible to micrococcal endonuclease digestion. The DNA fragments resulting from a mild endonuclease treatment are multiples of a basic unit of 194 +/- 6 base pairs in tobacco and 195 +/- 6 base pairs in barley. After extensive digestion, a DNA fragment of approximately...
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pub_type: 杂志文章
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更新日期:1977-10-01 00:00:00