Literature DB >> 9383474

Polymerase chain reaction in liposomes.

T Oberholzer1, M Albrizio, P L Luisi.   

Abstract

BACKGROUND: Compartmentalization of biochemical reactions within a spherically closed bilayer is an important step in the molecular evolution of cells. Liposomes are the most suitable structures to model this kind of chemistry. We have used the polymerase chain reaction (PCR) to demonstrate that complex biochemical reactions such as DNA replication can be carried out inside these compartments.
RESULTS: We describe the first example of DNA amplification by the PCR occurring inside liposomes composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), or of a mixture of POPC and phosphatidylserine. We show that these liposomes are stable even under the high temperature conditions used for PCR. Although only a very small fraction of liposomes contains all eight different reagents together, a significant amount of DNA is produced which can be observed by polyacrylamide gel electrophoresis.
CONCLUSIONS: This work shows that it is possible to carry out complex biochemical reactions within liposomes, which may be germane to the question of the origin of living cells. We have established the parameters and conditions that are critical for carrying out this complex reaction within the liposome compartment.

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Year:  1995        PMID: 9383474     DOI: 10.1016/1074-5521(95)90031-4

Source DB:  PubMed          Journal:  Chem Biol        ISSN: 1074-5521


  39 in total

1.  Nutrient uptake by protocells: a liposome model system.

Authors:  P A Monnard; D W Deamer
Journal:  Orig Life Evol Biosph       Date:  2001 Feb-Apr       Impact factor: 1.950

2.  Directed evolution of polymerase function by compartmentalized self-replication.

Authors:  F J Ghadessy; J L Ong; P Holliger
Journal:  Proc Natl Acad Sci U S A       Date:  2001-03-27       Impact factor: 11.205

3.  Self-reproduction of supramolecular giant vesicles combined with the amplification of encapsulated DNA.

Authors:  Kensuke Kurihara; Mieko Tamura; Koh-Ichiroh Shohda; Taro Toyota; Kentaro Suzuki; Tadashi Sugawara
Journal:  Nat Chem       Date:  2011-09-04       Impact factor: 24.427

4.  Targeted delivery of in situ PCR-amplified Sleeping Beauty transposon genes to cancer cells with lipid-based nanoparticle-like protocells.

Authors:  Kun Ma; Duo Fu; Dongli Yu; Changhao Cui; Li Wang; Zhaoming Guo; Chuanbin Mao
Journal:  Biomaterials       Date:  2017-01-02       Impact factor: 12.479

Review 5.  Approaches to semi-synthetic minimal cells: a review.

Authors:  Pier Luigi Luisi; Francesca Ferri; Pasquale Stano
Journal:  Naturwissenschaften       Date:  2006-01

6.  Piecing together a puzzle. An exposition of synthetic biology.

Authors:  Anna Deplazes
Journal:  EMBO Rep       Date:  2009-05       Impact factor: 8.807

7.  Models of primitive cellular life: polymerases and templates in liposomes.

Authors:  Pierre-Alain Monnard; Andrej Luptak; David W Deamer
Journal:  Philos Trans R Soc Lond B Biol Sci       Date:  2007-10-29       Impact factor: 6.237

8.  Stability of model membranes in extreme environments.

Authors:  Trishool Namani; David W Deamer
Journal:  Orig Life Evol Biosph       Date:  2008-06-17       Impact factor: 1.950

9.  Stochastic simulation and analysis of biomolecular reaction networks.

Authors:  John M Frazier; Yaroslav Chushak; Brent Foy
Journal:  BMC Syst Biol       Date:  2009-06-17

10.  Internal lipid synthesis and vesicle growth as a step toward self-reproduction of the minimal cell.

Authors:  Giovanni Murtas
Journal:  Syst Synth Biol       Date:  2009-12-03
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