Principle
The technique utilises the inherent biological activity of DNA topoisomerase I. The biological role of topoisomerase is to cleave and rejoin supercoiled DNA ends to facilitate replication. Vaccinia virus topoisomerase I specifically recognises DNA sequence 5ยด-(C/T)CCTT-3'. During replication, the enzyme digests DNA specifically at this sequence, unwinds the DNA and re-ligates it again at the 3' phosphate group of the thymidine base.
Read more about this topic: TOPO Cloning
Famous quotes containing the word principle:
“Thanks to all. For the great republic—for the principle it lives by, and keeps alive—for man’s vast future,—thanks to all.”
—Abraham Lincoln (1809–1865)
“To invent without scruple a new principle to every new phenomenon, instead of adapting it to the old; to overload our hypothesis with a variety of this kind, are certain proofs that none of these principles is the just one, and that we only desire, by a number of falsehoods, to cover our ignorance of the truth.”
—David Hume (1711–1776)
“Whether our feet are compressed in iron shoes, our faces hidden with veils and masks; whether yoked with cows to draw the plow through its furrows, or classed with idiots, lunatics and criminals in the laws and constitutions of the State, the principle is the same; for the humiliations of the spirit are as real as the visible badges of servitude.”
—Elizabeth Cady Stanton (1815–1902)