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Biology · 术语表

什么是 Restriction enzymes, ligase and vectors?

Definition 6.1 University Biology — Year 3 · 第 6 章 — Genetic Engineering and Biotechnology

A type II restriction enzyme is a bacterial endonuclease that cuts double-stranded DNA at a specific sequence of 4 to 84\text{ to }8 base pairs, usually a palindrome: EcoRI cuts G↓^{\downarrow}AATTC, leaving four-base single-stranded overhangs (sticky ends) complementary to any other EcoRI end; other enzymes leave blunt ends. DNA ligase seals two fragments whose ends fit. A vector is a DNA molecule that replicates in a host and can carry an inserted fragment: a plasmid of a few kilobases with an origin of replication, a selectable marker (an antibiotic resistance gene, so that only cells that took up the plasmid grow on the antibiotic) and a cluster of unique restriction sites for the insert; bacteriophage λ\lambda, cosmids, and bacterial or yeast artificial chromosomes for inserts of 20 to 1000 kb20\text{ to }1000\,\mathrm{kb}. DNA enters a bacterium by transformation, after a chemical or electrical shock that makes the membrane transiently permeable, with an efficiency of 10610^{6} to 10910^{9} transformants per microgram of plasmid. A recombinant molecule is one that joins DNA from two sources.

A cloning vector and its chemistry. The plasmid carries an origin, a selectable marker and a unique restriction site into which a fragment cut with the same enzyme is ligated; sticky ends make any two fragments cut by the same enzyme compatible.
A cloning vector and its chemistry. The plasmid carries an origin, a selectable marker and a unique restriction site into which a fragment cut with the same enzyme is ligated; sticky ends make any two fragments cut by the same enzyme compatible.
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