Biomaterial Database

Influence of nucleotide sequence adjacent to duplex DNA termini on 3' terminal labeling by terminal transferase. 1979

R Roychoudhury, and C P Tu, and R Wu

We have analyzed the effect of base-pairing at the exposed ends of Hinc II fragments of SV40 DNA on the efficiency of ribonucleotide incorporation catalyzed by terminal transferase. Wide variations in the labeling efficiency of individual DNA fragments have been observed. To elucidate the nature of this variation at the molecular level, we have correlated this effect with nucleotide sequence adjacent to the cleavage site. We found that a G:C base pair right at the exposed end drastically reduces the incorporation of ribonucleotides. Furthermore, the higher the number of G:C base pairs adjacent to the exposed end, the greater the reduction in labeling efficiency. From these results, we conclude that the labeling efficiency is determined by the degree of 'terminal breathing' of the DNA molecule at the exposed end of the duplex.

UI	MeSH Term	Description	Entries
D007700	Kinetics	The rate dynamics in chemical or physical systems.
D004254	DNA Nucleotidyltransferases	Enzymes that catalyze the incorporation of deoxyribonucleotides into a chain of DNA. EC 2.7.7.-.	Nucleotidyltransferases, DNA
D004262	DNA Restriction Enzymes	Enzymes that are part of the restriction-modification systems. They catalyze the endonucleolytic cleavage of DNA sequences which lack the species-specific methylation pattern in the host cell's DNA. Cleavage yields random or specific double-stranded fragments with terminal 5'-phosphates. The function of restriction enzymes is to destroy any foreign DNA that invades the host cell. Most have been studied in bacterial systems, but a few have been found in eukaryotic organisms. They are also used as tools for the systematic dissection and mapping of chromosomes, in the determination of base sequences of DNAs, and have made it possible to splice and recombine genes from one organism into the genome of another. EC 3.21.1.	Restriction Endonucleases,DNA Restriction Enzyme,Restriction Endonuclease,Endonuclease, Restriction,Endonucleases, Restriction,Enzymes, DNA Restriction,Restriction Enzyme, DNA,Restriction Enzymes, DNA
D004279	DNA, Viral	Deoxyribonucleic acid that makes up the genetic material of viruses.	Viral DNA
D001483	Base Sequence	The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.	DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA