PMID- 28815508
OWN - NLM
STAT- MEDLINE
DCOM- 20180426
LR  - 20230107
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 1642
DP  - 2017
TI  - Multipart DNA Assembly Using Site-Specific Recombinases from the Large Serine 
      Integrase Family.
PG  - 303-323
LID - 10.1007/978-1-4939-7169-5_19 [doi]
AB  - Assembling multiple DNA fragments into functional plasmids is an important and 
      often rate-limiting step in engineering new functions in living systems. 
      Bacteriophage integrases are enzymes that carry out efficient recombination 
      reactions between short, defined DNA sequences known as att sites. These DNA 
      splicing reactions can be used to assemble large numbers of DNA fragments into a 
      functional circular plasmid in a method termed serine integrase recombinational 
      assembly (SIRA). The resulting DNA assemblies can easily be modified by further 
      recombination reactions catalyzed by the same integrase in the presence of its 
      recombination directionality factor (RDF). Here we present a set of protocols for 
      the overexpression and purification of bacteriophage varphiC31 and Bxb1 integrase and 
      RDF proteins, their use in DNA assembly reactions, and subsequent modification of 
      the resulting DNA assemblies.
FAU - Olorunniji, Femi J
AU  - Olorunniji FJ
AD  - Institute of Molecular, Cell and Systems Biology, University of Glasgow, Bower 
      Building, Glasgow, G12 8QQ, Scotland.
FAU - Merrick, Christine
AU  - Merrick C
AD  - SynthSys-Synthetic and Systems Biology, School of Biological Sciences, University 
      of Edinburgh, CH Waddington Building, The King's Buildings, Mayfield Road, 
      Edinburgh, EH9 3JD, Scotland.
FAU - Rosser, Susan J
AU  - Rosser SJ
AD  - SynthSys-Synthetic and Systems Biology, School of Biological Sciences, University 
      of Edinburgh, CH Waddington Building, The King's Buildings, Mayfield Road, 
      Edinburgh, EH9 3JD, Scotland.
FAU - Smith, Margaret C M
AU  - Smith MCM
AD  - Department of Biology, University of York, Wentworth Way, York, YO10 5DD, UK.
FAU - Stark, W Marshall
AU  - Stark WM
AD  - Institute of Molecular, Cell and Systems Biology, University of Glasgow, Bower 
      Building, Glasgow, G12 8QQ, Scotland.
FAU - Colloms, Sean D
AU  - Colloms SD
AD  - Institute of Molecular, Cell and Systems Biology, University of Glasgow, Bower 
      Building, Glasgow, G12 8QQ, Scotland. Sean.Colloms@glasgow.ac.uk.
LA  - eng
GR  - BB/K011499/1/BB_/Biotechnology and Biological Sciences Research Council/United 
      Kingdom
GR  - BB/K011499/2/BB_/Biotechnology and Biological Sciences Research Council/United 
      Kingdom
GR  - BB/M018040/1/BB_/Biotechnology and Biological Sciences Research Council/United 
      Kingdom
PT  - Journal Article
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (DNA, Circular)
RN  - 0 (DNA, Viral)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Viral Proteins)
RN  - 452VLY9402 (Serine)
RN  - EC 2.7.7.- (DNA Nucleotidyltransferases)
RN  - EC 2.7.7.- (Integrases)
RN  - EC 2.7.7.- (Site-specific recombinase)
RN  - Streptomyces virus phiC31
SB  - IM
MH  - Attachment Sites, Microbiological
MH  - DNA Nucleotidyltransferases/*genetics/isolation & purification/metabolism
MH  - DNA, Circular/genetics/metabolism
MH  - DNA, Viral/genetics/metabolism
MH  - Escherichia coli/genetics/metabolism
MH  - Gene Expression
MH  - Integrases/*genetics/isolation & purification/metabolism
MH  - Metabolic Engineering/*methods
MH  - Plasmids/chemistry/*metabolism
MH  - Recombinant Proteins/genetics/isolation & purification/metabolism
MH  - Recombination, Genetic
MH  - Serine/metabolism
MH  - Siphoviridae/*genetics/metabolism
MH  - Viral Proteins/*genetics/isolation & purification/metabolism
OTO - NOTNLM
OT  - Bacteriophage varphiC31
OT  - DNA assembly
OT  - Large serine integrase
OT  - Metabolic engineering
OT  - Site-specific recombination
OT  - Synthetic biology
EDAT- 2017/08/18 06:00
MHDA- 2018/04/27 06:00
CRDT- 2017/08/18 06:00
PHST- 2017/08/18 06:00 [entrez]
PHST- 2017/08/18 06:00 [pubmed]
PHST- 2018/04/27 06:00 [medline]
AID - 10.1007/978-1-4939-7169-5_19 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2017;1642:303-323. doi: 10.1007/978-1-4939-7169-5_19.