Taq DNA ligase

DNA ligase
other names	Polydeoxyribonucleotide synthase [NAD (+)]
Mass / length primary structure	674 amino acids , 76,527 Da
Identifier
External IDs	UniProt : B7A6G7 ;
Enzyme classification
EC, category	6.5.1.2

The Taq DNA ligase is an enzyme from the group of DNA ligases and is produced by Thermus aquaticus .

properties

The Taq DNA ligase creates a phosphodiester bond between two adjacent DNA sequences in DNA double strands and is used by the bacteria for DNA replication , recombination and DNA repair . Cofactors are divalent magnesium ions .

The Taq DNA ligase catalyzes the reaction:

NAD ⁺ + (deoxyribonucleotide) (n) + (deoxyribonucleotide) (m) AMP + beta-nicotinamide-D-ribonucleotide + (deoxyribonucleotide) (n + m) ${\ displaystyle \ rightleftharpoons}$

Applications

The Taq DNA ligase is used in the course of an artificial gene synthesis for the multiple, parallel ligation of two DNA double strands to each other, but is not necessary when using the Gibson assembly for cloning . The Taq ligase is used in the course of a PCR optimization to generate long products, but independent of the ligase activity. Furthermore, the Taq DNA ligase is used for the ligase chain reaction .

Web links

MeSH Taq DNA ligase

Individual evidence

^ DG Gibson: Enzymatic assembly of overlapping DNA fragments. In: Methods in enzymology. Volume 498, 2011, pp. 349-361, doi : 10.1016 / B978-0-12-385120-8.00015-2 , PMID 21601685 .
^ RM Benoit, C. Ostermeier, M. Geiser, JS Li, H. Widmer, M. Auer: Seamless Insert-Plasmid Assembly at High Efficiency and Low Cost. In: PloS one. Volume 11, number 4, 2016, p. E0153158, doi : 10.1371 / journal.pone.0153158 , PMID 27073895 , PMC 4830597 (free full text).
↑ KB Ignatov, VM Kramarov: DNA ligases from thermophilic bacteria enhance PCR amplification of long DNA sequences. In: Biochemistry. Biokhimiia. Volume 74, Number 5, May 2009, pp. 557-561, PMID 19538130 .
↑ M. Wiedmann, WJ Wilson, J. Czajka, J. Luo, F. Barany, CA Batt: Ligase chain reaction (LCR) - review and applications. In: PCR methods and applications. Volume 3, Number 4, February 1994, pp. S51-S64, ISSN 1054-9803 . PMID 8173509 . (PDF).

[1] DG Gibson: Enzymatic assembly of overlapping DNA fragments. In: Methods in enzymology. Volume 498, 2011, pp. 349-361, doi : 10.1016 / B978-0-12-385120-8.00015-2 , PMID 21601685 .

[2] RM Benoit, C. Ostermeier, M. Geiser, JS Li, H. Widmer, M. Auer: Seamless Insert-Plasmid Assembly at High Efficiency and Low Cost. In: PloS one. Volume 11, number 4, 2016, p. E0153158, doi : 10.1371 / journal.pone.0153158 , PMID 27073895 , PMC 4830597 (free full text).

[3] KB Ignatov, VM Kramarov: DNA ligases from thermophilic bacteria enhance PCR amplification of long DNA sequences. In: Biochemistry. Biokhimiia. Volume 74, Number 5, May 2009, pp. 557-561, PMID 19538130 .

[4] M. Wiedmann, WJ Wilson, J. Czajka, J. Luo, F. Barany, CA Batt: Ligase chain reaction (LCR) - review and applications. In: PCR methods and applications. Volume 3, Number 4, February 1994, pp. S51-S64, ISSN 1054-9803 . PMID 8173509 . (PDF).