scholarly journals Insert + Vector DNA Ligation v1 (protocols.io.76uhrew)

protocols.io ◽  
2019 ◽  
Author(s):  
iGEM Dusseldorf
Keyword(s):  
Dna Ligation ◽  
Vector Dna

scholarly journals Efficient Transformation System forPropionibacterium freudenreichii Based on a Novel Vector

2001 ◽  
Vol 67 (2) ◽  
pp. 499-503 ◽  
Author(s):  
J. P. M. Jore ◽  
N. van Luijk ◽  
R. G. M. Luiten ◽  
M. J. van der Werf ◽  
P. H. Pouwels
Keyword(s):  
High Efficiency ◽  
Propionibacterium Freudenreichii ◽  
Transformation System ◽  
Erythromycin Resistance ◽  
E Coli ◽  
Recognition Sites ◽  
Dna Ligation ◽  
Enzyme Recognition ◽  
Restriction Modification ◽  
Vector Dna

ABSTRACT A 3.6-kb endogenous plasmid was isolated from aPropionibacterium freudenreichii strain and sequenced completely. Based on homologies with plasmids from other bacteria, notably a plasmid from Mycobacterium, a region harboring putative replicative functions was defined. Outside this region two restriction enzyme recognition sites were used for insertion of anEscherichia coli-specific replicon and an erythromycin resistance gene for selection in Propionibacterium. Hybrid vectors obtained in this way replicated in both E. coli andP. freudenreichii. Whereas electroporation of P. freudenreichii with vector DNA isolated from an E. coli transformant yielded 10 to 30 colonies per μg of DNA, use of vector DNA reisolated from a Propionibacteriumtransformant dramatically increased the efficiency of transformation (≥108 colonies per μg of DNA). It could be shown that restriction-modification was responsible for this effect. The high efficiency of the system described here permitted successful transformation of Propionibacterium with DNA ligation mixtures.

scholarly journals Structure-Function Analysis of IntDOT

2009 ◽  
Vol 192 (2) ◽  
pp. 575-586 ◽  
Author(s):  
Seyeun Kim ◽  
Brian M. Swalla ◽  
Jeffrey F. Gardner
Keyword(s):  
Homology Modeling ◽  
Protein Interactions ◽  
Active Site ◽  
Dna Cleavage ◽  
Function Analysis ◽  
Indicator Strain ◽  
Amino Terminal ◽  
Dna Ligation ◽  
Polypeptide Backbone

ABSTRACT CTnDOT integrase (IntDOT) is a member of the tyrosine family of site-specific DNA recombinases. IntDOT is unusual in that it catalyzes recombination between nonidentical sequences. Previous mutational analyses centered on mutants with substitutions of conserved residues in the catalytic (CAT) domain or residues predicted by homology modeling to be close to DNA in the core-binding (CB) domain. That work suggested that a conserved active-site residue (Arg I) of the CAT domain is missing and that some residues in the CB domain are involved in catalysis. Here we used a genetic approach and constructed an Escherichia coli indicator strain to screen for random mutations in IntDOT that disrupt integrative recombination in vivo. Twenty-five IntDOT mutants were isolated and characterized for DNA binding, DNA cleavage, and DNA ligation activities. We found that mutants with substitutions in the amino-terminal (N) domain were catalytically active but defective in forming nucleoprotein complexes, suggesting that they have altered protein-protein interactions or altered interactions with DNA. Replacement of Ala-352 of the CAT domain disrupted DNA cleavage but not DNA ligation, suggesting that Ala-352 may be important for positioning the catalytic tyrosine (Tyr-381) during cleavage. Interestingly, our biochemical data and homology modeling of the CAT domain suggest that Arg-285 is the missing Arg I residue of IntDOT. The predicted position of Arg-285 shows it entering the active site from a position on the polypeptide backbone that is not utilized in other tyrosine recombinases. IntDOT may therefore employ a novel active-site architecture to catalyze recombination.

Homologous and heterologous recombination between adenovirus vector DNA and chromosomal DNA

2008 ◽  
Vol 10 (11) ◽  
pp. 1176-1189 ◽  
Author(s):  
Sam Laurel Stephen ◽  
Vijayshankar Ganesh Sivanandam ◽  
Stefan Kochanek
Keyword(s):  
Adenovirus Vector ◽  
Chromosomal Dna ◽  
Vector Dna

scholarly journals Enzymes involved in DNA ligation and end-healing in the radioresistant bacterium Deinococcus radiodurans

2007 ◽  
Vol 8 (1) ◽  
pp. 69 ◽  
Author(s):  
Melanie Blasius ◽  
Rebecca Buob ◽  
Igor V Shevelev ◽  
Ulrich Hubscher
Keyword(s):  
Deinococcus Radiodurans ◽  
Dna Ligation

Adenoviral vector DNA for accurate genome editing with engineered nucleases

2014 ◽  
Vol 11 (10) ◽  
pp. 1051-1057 ◽  
Author(s):  
Maarten Holkers ◽  
Ignazio Maggio ◽  
Sara F D Henriques ◽  
Josephine M Janssen ◽  
Toni Cathomen ◽  
...  
Keyword(s):  
Genome Editing ◽  
Adenoviral Vector ◽  
Engineered Nucleases ◽  
Vector Dna

Label-free biosensor based on dsDNA-templated copper nanoparticles for highly sensitive and selective detection of NAD+

RSC Advances ◽  
2016 ◽  
Vol 6 (94) ◽  
pp. 91077-91082 ◽  
Author(s):  
Jia Ge ◽  
Zhen-Zhen Dong ◽  
Lin Zhang ◽  
Qi-Yong Cai ◽  
Dong-Mei Bai ◽  
...  
Keyword(s):  
Copper Nanoparticles ◽  
Selective Detection ◽  
Label Free ◽  
Ligation Reaction ◽  
Dna Ligation ◽  
Highly Sensitive

A novel label-free biosensor for high sensing of NAD+ based on dsDNA-templated CuNPs and DNA ligation reaction.

scholarly journals The DNA Repair Protein XRCC1 Functions in the Plant DNA Demethylation Pathway by Stimulating Cytosine Methylation (5-meC) Excision, Gap Tailoring, and DNA Ligation

2013 ◽  
Vol 288 (8) ◽  
pp. 5496-5505 ◽  
Author(s):  
María Isabel Martínez-Macías ◽  
Dolores Córdoba-Cañero ◽  
Rafael R. Ariza ◽  
Teresa Roldán-Arjona
Keyword(s):  
Dna Repair ◽  
Cytosine Methylation ◽  
Dna Demethylation ◽  
Plant Dna ◽  
Dna Ligation ◽  
Repair Protein ◽  
Demethylation Pathway ◽  
Dna Repair Protein
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