Enzymatic ligation

Another method is to enzymatically add suitable restriction sites to the blunt ends of the DNA. For this, a specially designed blunt-ended synthetic oligonucleotide species, known as a linker, is added and enzymatically ligated to the larger DNA. 

Preparation of the labeled constructs by enzymatic ligation involves several steps of preparing ligation components, as shown in Scheme 3.3. This protocol describes only the last step in the overall procedure, as the preceding steps are standard in RNA biochemistry and corresponding protocols can be found elsewhere (see, e.g., Akiyama and Stone, Chapter 2, this volume). 

Hobartner, C., and Micura, R. (2004). Chemical synthesis of selenium-modified oligo-ribonucleotides and their enzymatic ligation leading to an U6 SnRNA stem-loop segment. J. Am. Chem. Soc. 126, 1141—1149. 

Enzymatic Ligation of the Fragments Obtained by Site-selective Scission... 

Figure 7.7 Schematic representation of enzymatic ligation of the scission fragment and foreign DNA using various templates.

Figure 7.12 Agarose gel electrophoresis of the PCR product from the recombinant DNA. Lane 1, PCR product from the recombinant DNA lane 2, PCR product from the mixture without the enzymatic ligation M, 200 base pair ladder.

In the scission of double-stranded DNA by Ce(iv)/EDTA in the presence of pcPNA 1 /pcPNA 2, several fragments should be formed. In the enzymatic ligation, however, the scission fragment (fragment 1 here), whose sticky end completely fits... 

The development of EPL has facilitated the production of large protein targets, but the requirement of specific N-terminal amino acids at the ligation site (cysteine [7], selenocysteine ]66J) reduces the general utilization of this method. Recently, we introduced a novel approach that we named expressed enzymatic ligation (EEL) for the semisynthesis of larger and chemically modified proteins that combines the advantages of the EPL with those of the substrate mimetic... 

Machova, Z., von Eggelkraut-Gottanka, R., Wehofsky, N., Bordusa, E., and Beck-Sickinger, A. G. (2003) Expressed enzymatic ligation a new approach for the synthesis of chemically modified proteins, Angew. Chem. Int. Ed. 42, 4916-4918. 

In order to synthesize longer polypeptides and small proteins, the condensation of suitable starting fragments is an essential prerequisite. Since chemical fragment condensations suffer from the danger of enantiomerization, enzymatic ligations are pronoising alternatives (see... 

A hybrid dinucleotide pdCpA has been chemically aminoacylated with L-phenylalanine to give pdCpA-Phe. The aminoacylated dinucleotide can be used in an enzymatic ligation (T4 RNA llgase) and attached to the 3 -terminus of a truncated (3 -terminal CA residues missing) amber suppressor tRNA y. The resultant chemically aminoacylated tRNA can be used to incorporate L-phenylalanine in response to an amber nonsense coding. This procedure can... 

Fig. 9.8. Tetramers and hexamers of 10 nm Au nanocrystals assembled as chains of pairs of particles using DNA and enzymatic ligation. Collaborative work with Prof Jean Frechet [22]...

Frutos AG, Smith LM, Corn RM (1998) Enzymatic ligation reactions of DNA words on surfaces for DNA computing. J Am Chem Soc 120 10277-10282... 

Enzymatic ligation generates a crossed-strand intermediate, called a Holliday junction ... 

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