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Towards construction of synthetic ribosomes and a self-replicating system.

Record Type:	Language materials, manuscript : Monograph/item
Title/Author:	Towards construction of synthetic ribosomes and a self-replicating system./
Author:	Li, Jun.
Description:	1 online resource (104 pages)
Notes:	Source: Dissertation Abstracts International, Volume: 75-10(E), Section: B.
Contained By:	Dissertation Abstracts International75-10B(E).
Subject:	Biochemistry. -
Online resource:	click for full text (PQDT)
ISBN:	9781321018769

Towards construction of synthetic ribosomes and a self-replicating system.
Li, Jun.

Towards construction of synthetic ribosomes and a self-replicating system. - 1 online resource (104 pages)

Source: Dissertation Abstracts International, Volume: 75-10(E), Section: B.

Thesis (Ph.D.)

Includes bibliographical references

This item is not available from ProQuest Dissertations & Theses.

In 2006, the Church Group, using biochemical approaches, hypothesized that &sim;151 biomolecular components from Escherichia coli and its bacteriophages may be sufficient to enable rapid and accurate self-replication in vitro. However, efforts to construct such a system are precluded by our inability to sufficiently co-regenerate these 151 biomolecular components (or the components of ribosome---the key player of protein translation) and the inability to assemble E. coli ribosomes under conditions that are compatible with in vitro transcription and translation and also the lacking of evidence that functionally active ribosomes can be reconstituted from in vitro synthesized proteins and RNAs.

Electronic reproduction.
Ann Arbor, Mich. :
ProQuest,
2018

Mode of access: World Wide Web

ISBN: 9781321018769Subjects--Topical Terms:

582831
Biochemistry.
Index Terms--Genre/Form:

554714
Electronic books.

Towards construction of synthetic ribosomes and a self-replicating system.
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100 1 $a Li, Jun. $3 1073622
245 1 0 $a Towards construction of synthetic ribosomes and a self-replicating system.
264 0 $c 2014
300 $a 1 online resource (104 pages)
336 $a text $b txt $2 rdacontent
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500 $a Source: Dissertation Abstracts International, Volume: 75-10(E), Section: B.
500 $a Adviser: George M. Church.
502 $a Thesis (Ph.D.) $c Harvard University $d 2014.
504 $a Includes bibliographical references
506 $a This item is not available from ProQuest Dissertations & Theses.
520 $a In 2006, the Church Group, using biochemical approaches, hypothesized that &sim;151 biomolecular components from Escherichia coli and its bacteriophages may be sufficient to enable rapid and accurate self-replication in vitro. However, efforts to construct such a system are precluded by our inability to sufficiently co-regenerate these 151 biomolecular components (or the components of ribosome---the key player of protein translation) and the inability to assemble E. coli ribosomes under conditions that are compatible with in vitro transcription and translation and also the lacking of evidence that functionally active ribosomes can be reconstituted from in vitro synthesized proteins and RNAs.
520 $a In this study, I addressed these problems in three aspects. First, I selected a well-defined cell-free system--- Protein synthesis Using Recombinant Elements (PURE) system, as the platform to construct the self-replicating system and improved the PURE system productivity in the purpose of generating sufficient amount of protein for self-replicating. I added or adjusted the concentration of a variety of factors that affect transcription and translation and achieved a boost in PURE system productivity of more than 5 fold. The work provides a more efficient defined in vitro transcription and translation system and a deeper understanding of system efficiency limitation factors. Second, I demonstrated that the 54 E. coli ribosomal proteins can be individually expressed and co-regenerated in it. Third, in contrast to conventional non-physiological methods, I developed new methods for constructing E. coli ribosomes at physiological conditions by integrating ribosomal RNA modification, cofactor facilitated ribosome assembly and in vitro transcription and translation into a one-step incubation procedure. I also showed that fully synthetic functionally active 30S ribosome can be constructed from in vitro transcribed 16S rRNA and in vitro translated 30S ribosomal proteins. Our integrated strategy solved critical barriers to construct synthetic ribosomes and synthetic life and also pave the road to build a protein based self-replicating system.
533 $a Electronic reproduction. $b Ann Arbor, Mich. : $c ProQuest, $d 2018
538 $a Mode of access: World Wide Web
650 4 $a Biochemistry. $3 582831
650 4 $a Dentistry. $3 674038
655 7 $a Electronic books. $2 local $3 554714
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690 $a 0567
710 2 $a ProQuest Information and Learning Co. $3 1178819
710 2 $a Harvard University. $b Biological Sciences in Dental Medicine. $3 1184259
773 0 $t Dissertation Abstracts International $g 75-10B(E).
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3626824 $z click for full text (PQDT)