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Evolutionary principles applied to t...

Doyon, Jeffrey Brian.

Evolutionary principles applied to the discovery of small molecules, homing endonucleases, and eukaryotic promoter elements.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Evolutionary principles applied to the discovery of small molecules, homing endonucleases, and eukaryotic promoter elements.
作者:	Doyon, Jeffrey Brian.
面頁冊數:	144 p.
附註:	Adviser: David R. Liu.
附註:	Source: Dissertation Abstracts International, Volume: 67-05, Section: B, page: 2530.
Contained By:	Dissertation Abstracts International67-05B.
標題:	Chemistry, Biochemistry.
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3217716
ISBN:	9780542692338

Evolutionary principles applied to the discovery of small molecules, homing endonucleases, and eukaryotic promoter elements.
Doyon, Jeffrey Brian.

Evolutionary principles applied to the discovery of small molecules, homing endonucleases, and eukaryotic promoter elements. - 144 p.

Adviser: David R. Liu.

Thesis (Ph.D.)--Harvard University, 2006.

Chemists have traditionally used screens to discover small molecules with desired properties. In contrast, nature uses selections---processes by which molecules with favorable properties are physically separated from inactive molecules---to discover functional biological molecules. Chapter two describes the development of in vitro selections to identify DNA-linked small molecules with protein binding affinity and specificity. These selections require only generally accessible equipment, offer high degrees of enrichment of active molecules from mixtures of predominantly inactive species, can be applied to a variety of unrelated proteins, and require approximately 10 8 fold less material than existing synthetic molecule screening methods. The application of methods described in this work are currently playing a central role in the discovery of desired molecules from DNA-templated synthetic libraries.

ISBN: 9780542692338Subjects--Topical Terms:

226900
Chemistry, Biochemistry.

Evolutionary principles applied to the discovery of small molecules, homing endonucleases, and eukaryotic promoter elements.
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520 # $a Chemists have traditionally used screens to discover small molecules with desired properties. In contrast, nature uses selections---processes by which molecules with favorable properties are physically separated from inactive molecules---to discover functional biological molecules. Chapter two describes the development of in vitro selections to identify DNA-linked small molecules with protein binding affinity and specificity. These selections require only generally accessible equipment, offer high degrees of enrichment of active molecules from mixtures of predominantly inactive species, can be applied to a variety of unrelated proteins, and require approximately 10 8 fold less material than existing synthetic molecule screening methods. The application of methods described in this work are currently playing a central role in the discovery of desired molecules from DNA-templated synthetic libraries.
520 # $a Researchers have drawn inspiration from the wide variety of diversification methods used by nature during evolution to generate complex DNA libraries for directed evolution experiments. Our group has previously reported a method called nonhomologous random recombination (NRR) that allows for the extensive rearrangement and minimization of nucleic acids with no requirement for sequence homology between rearranged DNA segments. Chapter four describes the functional dissection of yeast promoters involved in the unfolded protein response using NRR diversification coupled with in vivo screening. This dissection rapidly identified new cis-regulatory elements in promoters implicated in the unfolded protein response.
520 # $a The laboratory evolution of enzymes with tailor-made DNA cleavage specificities would represent new tools for manipulating genomes. Chapter three describes the development of an efficient in vivo positive and negative selection system that applies evolutionary pressure either to favor the cleavage of a desired target sequence or to disfavor the cleavage of non-target sequences. We also applied a previously described in vitro selection method to reveal the comprehensive substrate specificity profile of the wild-type I-SceI homing endonuclease. Together these tools were applied to successfully evolve mutant I-SceI homing endonucleases with altered DNA cleavage specificities.
520 # $a This thesis describes three projects in which we have applied principles underlying biological evolution to evaluate small molecules, create new homing endonuclease enzymes, and functionally dissect yeast promoters.
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