Terminal deoxynucleotidyltransferase

Delarue, M., Boule, J. B., Lescar, J., Expert-Bezancon, N., Jourdan, N., Sukumar, N., Rougeon, R, and Papanicolaou, C. (2002) Crystal structures of a template-independent DNA polymerase murine terminal deoxynucleotidyltransferase. EMBO J. 21,427-439. 

DNA damage was evaluated by the terminal deoxynucleotidyltransferase (TdT)-mediated UTP nick end labeling (TUNEL) assay. Degenerating cells were stained by the Fluoro-Jade dye. 

Using terminal deoxynucleotidyltransferase -1- [a- P]ddATP, a 36-nucleotide oli-godeoxyribonucleotide containing uridine at position 21 is labeled at its 3 -end and the product is subjected to 20% denaturing polyacrylamide gel electrophoresis for... 

The extension of a polymer chain through enzymatic polymerisation results in a material response when the self-assembling properties of the polymer are changed as a result. Phosphorylase b has been used to extend the amylose part of a polymer amphiphUe by attaching further o-glucose-l-phosphate units to the end of the amylose chain (Morimoto et al., 2007) (Fig. 6.13). Terminal deoxynucleotidyltransferase (a DNA polymerase) was used to... 

In contrast to the DNA polymerases that require templates, some DNA polymerases conventionally known as template-independent DNA polymerases (e.g., terminal deoxynucleotidyltransferases) catalyze polymerization reactions in vitro without a template in the presence of dNTPs and an initiator (3 -OH). [Note In the enzyme classifications recommended by lUBMB, DNA polymerases that require a template and a primer belong to a subclass oi nucleotidyltransferases (EC 2.7.7).]... 

The third alternative is the homopolymeric tailing of cDNA using terminal deoxynucleotidyltransferase and annealing with the vector carrying complementary homopolymeric tails (see Section IV, this chapter). 

Terminal deoxynucleotidyltransferase is a group of DNA polymerases which, unlike DNA- or RNA-directed polymerases, require neither DNA or RNA templates but catalyze linear condensation polymerization reactions. Terminal deoxynucleotidyltransferase is traditionally abbreviated as TdT, However, this book will break with the tradition and use TDTase to avoid any possible confusion with the dinucleotide TdT. TDTase, which was first isolated from the calf thymus gland, is found almost exclusively in the thymuses of a wide variety of mammals and birds and in certain lymphoid cells. TDTase from calf thymus is a heterodimer (44 kDa) and both subunits are derived from a single polypeptide precursor ( 60 kDa). 

Isotopic labeling of DNA (and RNA as well) at the 3 or 5 end is typically performed using polynucleotide kinase and [y- P] ATP (refer to Section II, Chapter 5). Some 3 -end labeling can also be performed with DNA polymerase or terminal deoxynucleotidyltransferase and [ - P]dNTP or [a- P]ddNTP. Labeling by incorporation of isotopically or nonisotopically labeled nucleotides usually relies on DNA (or RNA) polymerases, although nonisotopically labeled oligonucleotides... 

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